Plant sugar transport proteins

ABSTRACT

This invention relates to an isolated nucleic acid fragment encoding a sugar transport protein. The invention also relates to the construction of a chimeric gene encoding all or a portion of the sugar transport protein, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the sugar transport protein in a transformed host cell.

This application claims the benefit of U.S. Provisional Application 60/083,044, filed Apr. 24, 1998.

FIELD OF THE INVENTION

This invention is in the field of plant molecular biology. More specifically, this invention pertains to nucleic acid fragments encoding sugar transport proteins in plants and seeds.

BACKGROUND OF THE INVENTION

Sugar is one form of carbohydrate produced in photosynthesizing cells in most higher plants and is the main form of transported carbon in most annual field crops such as corn, rice, soybeans and wheat. As such its movement and concentration across various plant membranes is critical to plant growth and development. In addition sugar is the main form of carbon that moves into developing seeds of soybeans, rice, corn and wheat. This movement and concentration is accomplished by the action of carrier proteins that act to transport sugar against a concentration gradient often by coupling sugar movement to the opposite vectoral movement of a proton. Specific sugar carrier proteins from these crop plants could be manipulated in efforts to control carbon flux and the timing and extent of sugar transport phenomena (e.g., grain fill duration) that are important factors in crop yield and quality. Accordingly, the availability of nucleic acid sequences encoding all or a portion of sugar transport proteins would facilitate studies to better understand carbon flux and sugar transport in plants, provide genetic tools for the manipulation of sugar transport, and provide a means to control carbohydrate transport and distribution in plant cells.

SUMMARY OF THE INVENTION

The instant invention relates to isolated nucleic acid fragments encoding sugar transport proteins. Specifically, this invention concerns an isolated nucleic acid fragment encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein. In addition, this invention relates to a nucleic acid fragment that is complementary to the nucleic acid fragment encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein.

An additional embodiment of the instant invention pertains to a polypeptide encoding all or a substantial portion of a sugar transport protein selected from the group consisting of Arabidopsis thaliana-like sugar transport protein and Beta vulgaris-like sugar transport protein.

In another embodiment, the instant invention relates to a chimeric gene encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein, or to a chimeric gene that comprises a nucleic acid fragment that is complementary to a nucleic acid fragment encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein, operably linked to suitable regulatory sequences, wherein expression of the chimeric gene results in production of levels of the encoded protein in a transformed host cell that is altered (i.e., increased or decreased) from the level produced in an untransformed host cell.

In a further embodiment, the instant invention concerns a transformed host cell comprising in its genome a chimeric gene encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein, operably linked to suitable regulatory sequences. Expression of the chimeric gene results in production of altered levels of the encoded protein in the transformed host cell. The transformed host cell can be of eukaryotic or prokaryotic origin, and include cells derived from higher plants and microorganisms. The invention also includes transformed plants that arise from transformed host cells of higher plants, and seeds derived from such transformed plants.

An additional embodiment of the instant invention concerns a method of altering the level of expression of an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein in a transformed host cell comprising: a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein; and b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of altered levels of Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein in the transformed host cell.

An additional embodiment of the instant invention concerns a method for obtaining a nucleic acid fragment encoding all or a substantial portion of an amino acid sequence encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein.

BRIEF DESCRIPTION OF THE DRAWINGS AND SEQUENCE DESCRIPTIONS

The invention can be more fully understood from the following detailed description and the accompanying drawings and Sequence Listing which form a part of this application.

FIGS. 1A-1E show a comparison of the amino acid sequences set forth in SEQ ID NOS:2, 4, 6, 8, 10, 12, 14 and 16 with the Arabidopsis thaliana-like sugar transport protein amino acid sequence set forth in SEQ ID NO:29. Amino acid designations in small case letters represent regions that are thought to be Arabidopsis thaliana-like sugar transport protein signatures.

FIG. 2 shows a comparison of the amino acid sequences set forth in SEQ ID NOS:18, 35 20, 22, 24, 26 and 28 with the Beta vulgaris-like sugar transport protein amino acid sequence set forth in SEQ ID NO:30.

The following sequence descriptions and Sequence Listing attached hereto comply with the rules governing nucleotide and/or amino acid sequence disclosures in patent applications as set forth in 37 C.F.R. §1.821-1.825.

SEQ ID NO:1 is the nucleotide sequence comprising a contig assembled from the DNA inserts in clones p0032.crcba66r, p0097.cqran41r, cr1n.pk0143.h10, p0128.cpict38, p0106.cjlpm67r, cil1c.pk001.f21, p0072.comgi92r, p0114.cimm181r and p0002.cgevb73r encoding a corn Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:2 is the deduced amino acid sequence of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO: 1.

SEQ ID NO:3 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones rlr12.pk0013.d11 and rds1c.pk007.n17 encoding a portion of a rice Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:4 is the deduced amino acid sequence of a portion of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:3.

SEQ ID NO:5 is the nucleotide sequence comprising a the entire cDNA insert in clone rls6.pk0003.d5 encoding a portion of a rice Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:6 is the deduced amino acid sequence of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:5.

SEQ ID NO:7 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones sgs4c.pk005.c9, sfl1.pk0079.a4 and sdp3c.pk012.i1 encoding a soybean Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:8 is the deduced amino acid sequence of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:7.

SEQ ID NO:9 is the nucleotide sequence comprising a portion of the cDNA insert in clone ss1.pk0022.f1 encoding a portion of a soybean Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:10 is the deduced amino acid sequence of a portion of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:9.

SEQ ID NO:11 is the nucleotide sequence comprising a portion of the cDNA insert in clone wlk8.pk0001.a12 encoding a portion of a wheat Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:12 is the deduced amino acid sequence of a portion of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:11.

SEQ ID NO:13 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones wlm96.pk043.e19 and wre1n.pk0062.g6 encoding a portion of a wheat Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:14 is the deduced amino acid sequence of a portion of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:13.

SEQ ID NO:15 is the nucleotide sequence comprising a portion of the cDNA insert in clone wre1n.pk0006.b4 encoding a portion of a wheat Arabidopsis thaliana-like sugar transport protein.

SEQ ID NO:16 is the deduced amino acid sequence of a portion of an Arabidopsis thaliana-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:15.

SEQ ID NO:17 is the nucleotide sequence comprising a portion of the cDNA insert in clone cc1.mn0002.h1 encoding a portion of a corn Beta vulgaris-like sugar transport protein.

SEQ ID NO:18 is the deduced amino acid sequence of a portion of a Beta vulgaris-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:17.

SEQ ID NO:19 is the nucleotide sequence comprising the entire cDNA insert in clone cepe7.pk0018.g3 encoding a corn Beta vulgaris-like sugar transport protein.

SEQ ID NO:20 is the deduced amino acid sequence of a Beta vulgaris-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:19.

SEQ ID NO:21 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones rlr6.pk0005.b10, r10n.pk102.p24 and r10n.pk107.p2 encoding a rice Beta vulgaris-like sugar transport protein.

SEQ ID NO:22 is the deduced amino acid sequence of a Beta vulgaris-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:21.

SEQ ID NO:23 is the nucleotide sequence comprising a contig assembled from the cDNA inserts in clones sr1.pk0061.g8, sfl1.pk0058.h12, sgs2c.pk004.o17 and sre.pk0032.h6 encoding a soybean Beta vulgaris-like sugar transport protein.

SEQ ID NO:24 is the deduced amino acid sequence of a Beta vulgaris-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:23.

SEQ ID NO:25 is the nucleotide sequence comprising the entire cDNA insert in clone wlk8.pk0001.a 11 encoding a wheat Beta vulgaris-like sugar transport protein.

SEQ ID NO:26 is the deduced amino acid sequence of a Beta vulgaris-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:25.

SEQ ID NO:27 is the nucleotide sequence comprising the entire cDNA insert in clone wlm1.pk0012.h1 encoding a wheat Beta vulgaris-like sugar transport protein.

SEQ ID NO:28 is the deduced amino acid sequence of a Beta vulgaris-like sugar transport protein derived from the nucleotide sequence of SEQ ID NO:28.

SEQ ID NO:29 is the amino acid sequence of an Arabidopsis thaliana (NCBI Identification No. gi 3080420) sugar transport protein.

SEQ ID NO:30 is the amino acid sequence of a Beta vulgaris (NCBI Identification No. gi 1778093) sugar transport protein.

The Sequence Listing contains the one letter code for nucleotide sequence characters and the three letter codes for amino acids as defined in conformity with the IUPAC-IUBMB standards described in Nucleic Acids Research 13:3021-3030 (1985) and in the Biochemical Journal 219 (No. 2):345-373 (1984) which are herein incorporated by reference. The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. §1.822.

DETAILED DESCRIPTION OF THE INVENTION

In the context of this disclosure, a number of terms shall be utilized. As used herein, an “isolated nucleic acid fragment” is a polymer of RNA or DNA that is single- or double-stranded, optionally containing synthetic, non-natural or altered nucleotide bases. An isolated nucleic acid fragment in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA or synthetic DNA. As used herein, “contig” refers to an assemblage of overlapping nucleic acid sequences to form one contiguous nucleotide sequence. For example, several DNA sequences can be compared and aligned to identify common or overlapping regions. The individual sequences can then be assembled into a single contiguous nucleotide sequence.

As used herein, “substantially similar” refers to nucleic acid fragments wherein changes in one or more nucleotide bases results in substitution of one or more amino acids, but do not affect the functional properties of the protein encoded by the DNA sequence. “Substantially similar” also refers to nucleic acid fragments wherein changes in one or more nucleotide bases does not affect the ability of the nucleic acid fragment to mediate alteration of gene expression by antisense or co-suppression technology. “Substantially similar” also refers to modifications of the nucleic acid fragments of the instant invention such as deletion or insertion of one or more nucleotides that do not substantially affect the functional properties of the resulting transcript vis-à-vis the ability to mediate alteration of gene expression by antisense or co-suppression technology or alteration of the functional properties of the resulting protein molecule. It is therefore understood that the invention encompasses more than the specific exemplary sequences.

For example, it is well known in the art that antisense suppression and co-suppression of gene expression may be accomplished using nucleic acid fragments representing less than the entire coding region of a gene, and by nucleic acid fragments that do not share 100% sequence identity with the gene to be suppressed. Moreover, alterations in a gene which result in the production of a chemically equivalent amino acid at a given site, but do not effect the functional properties of the encoded protein, are well known in the art. Thus, a codon for the amino acid alanine, a hydrophobic amino acid, may be substituted by a codon encoding another less hydrophobic residue, such as glycine, or a more hydrophobic residue, such as valine, leucine, or isoleucine. Similarly, changes which result in substitution of one negatively charged residue for another, such as aspartic acid for glutamic acid, or one positively charged residue for another, such as lysine for arginine, can also be expected to produce a functionally equivalent product. Nucleotide changes which result in alteration of the N-terminal and C-terminal portions of the protein molecule would also not be expected to alter the activity of the protein. Each of the proposed modifications is well within the routine skill in the art, as is determination of retention of biological activity of the encoded products.

Moreover, substantially similar nucleic acid fragments may also be characterized by their ability to hybridize, under stringent conditions (0.1×SSC, 0.1% SDS, 65° C.), with the nucleic acid fragments disclosed herein.

Substantially similar nucleic acid fragments of the instant invention may also be characterized by the percent similarity of the amino acid sequences that they encode to the amino acid sequences disclosed herein, as determined by algorithms commonly employed by those skilled in this art. Preferred are those nucleic acid fragments whose nucleotide sequences encode amino acid sequences that are 90% similar to the amino acid sequences reported herein. Most preferred are nucleic acid fragments that encode amino acid sequences that are 95% similar to the amino acid sequences reported herein. Sequence alignments and percent similarity calculations were performed using the Megalign program of the LASARGENE bioinformatics computing suite (DNASTAR Inc., Madison, Wis.). Multiple alignment of the sequences was performed using the Clustal method of alignment (Higgins, D. G. and Sharp, P. M. (1989) CABIOS. 5:151-153) with the default parameters (GAP PENALTY=10, GAP LENGTH PENALTY=10) (hereafter, Clustal algorithm). Default parameters for pairwise alignments using the Clustal method were KTUPLE 1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5.

A “substantial portion” of an amino acid or nucleotide sequence comprises enough of the amino acid sequence of a polypeptide or the nucleotide sequence of a gene to afford putative identification of that polypeptide or gene, either by manual evaluation of the sequence by one skilled in the art, or by computer-automated sequence comparison and identification using algorithms such as BLAST (Basic Local Alignment Search Tool; Altschul, S. F., et al., (1993) J. Mol. Biol. 215:403-410; see also www.ncbi.nlm.nih.gov/BLAST/). In general, a sequence of ten or more contiguous amino acids or thirty or more nucleotides is necessary in order to putatively identify a polypeptide or nucleic acid sequence as homologous to a known protein or gene. Moreover, with respect to nucleotide sequences, gene specific oligonucleotide probes comprising 20-30 contiguous nucleotides may be used in sequence-dependent methods of gene identification (e.g., Southern hybridization) and isolation (e.g., in situ hybridization of bacterial colonies or bacteriophage plaques). In addition, short oligonucleotides of 12-15 bases may be used as amplification primers in PCR in order to obtain a particular nucleic acid fragment comprising the primers. Accordingly, a “substantial portion” of a nucleotide sequence comprises enough of the sequence to afford specific identification and/or isolation of a nucleic acid fragment comprising the sequence. The instant specification teaches partial or complete amino acid and nucleotide sequences encoding one or more particular plant proteins. The skilled artisan, having the benefit of the sequences as reported herein, may now use all or a substantial portion of the disclosed sequences for purposes known to those skilled in this art. Accordingly, the instant invention comprises the complete sequences as reported in the accompanying Sequence Listing, as well as substantial portions of those sequences as defined above.

“Codon degeneracy” refers to divergence in the genetic code permitting variation of the nucleotide sequence without effecting the amino acid sequence of an encoded polypeptide. Accordingly, the instant invention relates to any nucleic acid fragment that encodes all or a substantial portion of the amino acid sequence encoding the Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins as set forth in SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26 and 28. The skilled artisan is well aware of the “codon-bias” exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid. Therefore, when synthesizing a gene for improved expression in a host cell, it is desirable to design the gene such that its frequency of codon usage approaches the frequency of preferred codon usage of the host cell.

“Synthetic genes” can be assembled from oligonucleotide building blocks that are chemically synthesized using procedures known to those skilled in the art. These building blocks are ligated and annealed to form gene segments which are then enzymatically assembled to construct the entire gene. “Chemically synthesized”, as related to a sequence of DNA, means that the component nucleotides were assembled in vitro. Manual chemical synthesis of DNA may be accomplished using well established procedures, or automated chemical synthesis can be performed using one of a number of commercially available machines. Accordingly, the genes can be tailored for optimal gene expression based on optimization of nucleotide sequence to reflect the codon bias of the host cell. The skilled artisan appreciates the likelihood of successful gene expression if codon usage is biased towards those codons favored by the host. Determination of preferred codons can be based on a survey of genes derived from the host cell where sequence information is available.

“Gene” refers to a nucleic acid fragment that expresses a specific protein, including regulatory sequences preceding (5′ non-coding sequences) and following (3′ non-coding sequences) the coding sequence. “Native gene” refers to a gene as found in nature with its own regulatory sequences. “Chimeric gene” refers any gene that is not a native gene, comprising regulatory and coding sequences that are not found together in nature. Accordingly, a chimeric gene may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different than that found in nature. “Endogenous gene” refers to a native gene in its natural location in the genome of an organism. A “foreign” gene refers to a gene not normally found in the host organism, but that is introduced into the host organism by gene transfer. Foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes. A “transgene” is a gene that has been introduced into the genome by a transformation procedure.

“Coding sequence” refers to a DNA sequence that codes for a specific amino acid sequence. “Regulatory sequences” refer to nucleotide sequences located upstream (5′ non-coding sequences), within, or downstream (3′ non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include promoters, translation leader sequences, introns, and polyadenylation recognition sequences. “Promoter” refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA. In general, a coding sequence is located 3′ to a promoter sequence. The promoter sequence consists of proximal and more distal upstream elements, the latter elements often referred to as enhancers. Accordingly, an “enhancer” is a DNA sequence which can stimulate promoter activity and may be an innate element of the promoter or a heterologous element inserted to enhance the level or tissue-specificity of a promoter. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. Promoters which cause a gene to be expressed in most cell types at most times are commonly referred to as “constitutive promoters”. New promoters of various types useful in plant cells are constantly being discovered; numerous examples may be found in the compilation by Okamuro and Goldberg, (1989) Biochemistry of Plants 15:1-82. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of different lengths may have identical promoter activity.

The “translation leader sequence” refers to a DNA sequence located between the promoter sequence of a gene and the coding sequence. The translation leader sequence is present in the fully processed MRNA upstream of the translation start sequence. The translation leader sequence may affect processing of the primary transcript to MRNA, mRNA stability or translation efficiency. Examples of translation leader sequences have been described (Turner, R. and Foster, G. D. (1995) Molecular Biotechnology 3:225).

The “3′ non-coding sequences” refer to DNA sequences located downstream of a coding sequence and include polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal is usually characterized by affecting the addition of polyadenylic acid tracts to the 3′ end of the mRNA precursor. The use of different 3′ non-coding sequences is exemplified by Ingelbrecht et al., (1989) Plant Cell 1:671-680.

“RNA transcript” refers to the product resulting from RNA polymerase-catalyzed transcription of a DNA sequence. When the RNA transcript is a perfect complementary copy of the DNA sequence, it is referred to as the primary transcript or it may be a RNA sequence derived from posttranscriptional processing of the primary transcript and is referred to as the mature RNA. “Messenger RNA (mRNA)” refers to the RNA that is without introns and that can be translated into protein by the cell. “cDNA” refers to a double-stranded DNA that is complementary to and derived from mRNA. “Sense” RNA refers to RNA transcript that includes the mRNA and so can be translated into protein by the cell. “Antisense RNA” refers to a RNA transcript that is complementary to all or part of a target primary transcript or MRNA and that blocks the expression of a target gene (U.S. Pat. No. 5,107,065, incorporated herein by reference). The complementarity of an antisense RNA may be with any part of the specific gene transcript, i.e., at the 5′ non-coding sequence, 3′ non-coding sequence, introns, or the coding sequence. “Functional RNA” refers to sense RNA, antisense RNA, ribozyme RNA, or other RNA that may not be translated but yet has an effect on cellular processes.

The term “operably linked” refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably linked with a coding sequence when it is capable of affecting the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory sequences in sense or antisense orientation.

The term “expression”, as used herein, refers to the transcription and stable accumulation of sense (mRNA) or antisense RNA derived from the nucleic acid fragment of the invention. Expression may also refer to translation of mRNA into a polypeptide. “Antisense inhibition” refers to the production of antisense RNA transcripts capable of suppressing the expression of the target protein. “Overexpression” refers to the production of a gene product in transgenic organisms that exceeds levels of production in normal or non-transformed organisms. “Co-suppression” refers to the production of sense RNA transcripts capable of suppressing the expression of identical or substantially similar foreign or endogenous genes (U.S. Pat. No. 5,231,020, incorporated herein by reference). “Altered levels” refers to the production of gene product(s) in transgenic organisms in amounts or proportions that differ from that of normal or non-transformed organisms.

“Mature” protein refers to a post-translationally processed polypeptide; i.e., one from which any pre- or propeptides present in the primary translation product have been removed. “Precursor” protein refers to the primary product of translation of mRNA; i.e., with pre- and propeptides still present. Pre- and propeptides may be but are not limited to intracellular localization signals.

A “chloroplast transit peptide” is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the chloroplast or other plastid types present in the cell in which the protein is made. “Chloroplast transit sequence” refers to a nucleotide sequence that encodes a chloroplast transit peptide. A “signal peptide” is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the secretory system (Chrispeels, J. J., (1991) Ann. Rev. Plant Phys. Plant Mol. Biol. 42:21-53). If the protein is to be directed to a vacuole, a vacuolar targeting signal (supra) can further be added, or if to the endoplasmic reticulum, an endoplasmic reticulum retention signal (supra) may be added. If the protein is to be directed to the nucleus, any signal peptide present should be removed and instead a nuclear localization signal included (Raikhel (1992) Plant Phys. 100:1627-1632).

“Transformation” refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in genetically stable inheritance. Host organisms containing the transformed nucleic acid fragments are referred to as “transgenic” organisms. Examples of methods of plant transformation include Agrobacterium-mediated transformation (De Blaere et al. (1987) Meth. Enzymol. 143:277) and particle-accelerated or “gene gun” transformation technology (Klein et al. (1987) Nature (London) 327:70-73; U.S. Pat. No. 4,945,050, incorporated herein by reference).

Standard recombinant DNA and molecular cloning techniques used herein are well known in the art and are described more fully in Sambrook, J., Fritsch, E. F. and Maniatis, T. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, 1989 (hereinafter “Maniatis”).

Nucleic acid fragments encoding at least a portion of several sugar transport proteins have been isolated and identified by comparison of random plant cDNA sequences to public databases containing nucleotide and protein sequences using the BLAST algorithms well known to those skilled in the art. Table 1 lists the proteins that are described herein, and the designation of the cDNA clones that comprise the nucleic acid fragments encoding these proteins.

TABLE 1 Sugar Transport Proteins Enzyme Clone Plant Sugar Transport Protein (Arabidopsis-like) p0032.crcba66r Corn p0097.cqran41r Corn crln.pk0143.h10 Corn p0128.cpict38 Corn p0106.cjlpm67r Corn ci1lc.pk001.f21 Corn p0072.comgi92r Corn p0114.cimm181r Corn p0002.cgevb73r Corn rdslc.pk007.n17 Rice rlr12.pk0013.d11 Rice rls6.pk0003.d5 Rice sgs4c.pk005.c9 Soybean sfl1.pk0079.a4 Soybean sdp3c.pk012.i1 Soybean ss1.pk0022.f1 Soybean wlk8.pk0001.a12 Wheat wlm96.pk043.e19 Wheat wre1n.pk0062.g6 Wheat wre1n.pk0006.b4 Wheat Sugar Transport Protein cc1.mn0002.h1 Corn (Beta vulgaris-like) cepe7.pk0018.g3 Corn rlr6.pk0005.b10 Rice rl0n.pk102.p24 Rice rl0n.pk107.p2 Rice sr1.pk0061.g8 Soybean sfl1.pk0058.h12 Soybean sgs2c.pk004.o17 Soybean sre.pk0032.h6 Soybean wlk8.pk0001.a11 Wheat wlml.pk0012.h1 Wheat

The nucleic acid fragments of the instant invention may be used to isolate cDNAs and genes encoding homologous proteins from the same or other plant species. Isolation of homologous genes using sequence-dependent protocols is well known in the art. Examples of sequence-dependent protocols include, but are not limited to, methods of nucleic acid hybridization, and methods of DNA and RNA amplification as exemplified by various uses of nucleic acid amplification technologies (e.g., polymerase chain reaction, ligase chain reaction).

For example, genes encoding other Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins, either as cDNAs or genomic DNAs, could be isolated directly by using all or a portion of the instant nucleic acid fragments as DNA hybridization probes to screen libraries from any desired plant employing methodology well known to those skilled in the art. Specific oligonucleotide probes based upon the instant nucleic acid sequences can be designed and synthesized by methods known in the art (Maniatis). Moreover, the entire sequences can be used directly to synthesize DNA probes by methods known to the skilled artisan such as random primer DNA labeling, nick translation, or end-labeling techniques, or RNA probes using available in vitro transcription systems. In addition, specific primers can be designed and used to amplify a part or all of the instant sequences. The resulting amplification products can be labeled directly during amplification reactions or labeled after amplification reactions, and used as probes to isolate fall length cDNA or genomic fragments under conditions of appropriate stringency.

In addition, two short segments of the instant nucleic acid fragments may be used in polymerase chain reaction protocols to amplify longer nucleic acid fragments encoding homologous genes from DNA or RNA. The polymerase chain reaction may also be performed on a library of cloned nucleic acid fragments wherein the sequence of one primer is derived from the instant nucleic acid fragments, and the sequence of the other primer takes advantage of the presence of the polyadenylic acid tracts to the 3′ end of the MnRNA precursor encoding plant genes. Alternatively, the second primer sequence may be based upon sequences derived from the cloning vector. For example, the skilled artisan can follow the RACE protocol (Frohman et al., (1988) PNAS USA 85:8998) to generate cDNAs by using PCR to amplify copies of the region between a single point in the transcript and the 3′ or 5′ end. Primers oriented in the 3′ and 5′ directions can be designed from the instant sequences. Using commercially available 3′ RACE or 5′ RACE systems (BRL), specific 3′ or 5′ cDNA fragments can be isolated (Ohara et al., (1989) PNAS USA 86:5673; Loh et al., (1989) Science 243:217). Products generated by the 3′ and 5′ RACE procedures can be combined to generate full-length cDNAs (Frohman, M. A. and Martin, G. R., (1989) Techniques 1:165).

Availability of the instant nucleotide and deduced amino acid sequences facilitates immunological screening of cDNA expression libraries. Synthetic peptides representing portions of the instant amino acid sequences may be synthesized. These peptides can be used to immunize animals to produce polyclonal or monoclonal antibodies with specificity for peptides or proteins comprising the amino acid sequences. These antibodies can be then be used to screen cDNA expression libraries to isolate full-length cDNA clones of interest (Lemer, R. A. (1984) Adv. Immunol. 36:1; Maniatis).

The nucleic acid fragments of the instant invention may be used to create transgenic plants in which the disclosed Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins are present at higher or lower levels than normal or in cell types or developmental stages in which they are not normally found. This would have the effect of altering the level of sugar transport in those cells.

Overexpression of the Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins of the instant invention may be accomplished by first constructing a chimeric gene in which the coding region is operably linked to a promoter capable of directing expression of a gene in the desired tissues at the desired stage of 35 development. For reasons of convenience, the chimeric gene may comprise promoter sequences and translation leader sequences derived from the same genes. 3′ Non-coding sequences encoding transcription termination signals may also be provided. The instant chimeric gene may also comprise one or more introns in order to facilitate gene expression.

Plasmid vectors comprising the instant chimeric gene can then constructed. The choice of plasmid vector is dependent upon the method that will be used to transform host plants. The skilled artisan is well aware of the genetic elements that must be present on the plasmid vector in order to successfully transform, select and propagate host cells containing the chimeric gene. The skilled artisan will also recognize that different independent transformation events will result in different levels and patterns of expression (Jones et al., (1985) EMBO J. 4:2411-2418; De Almeida et al., (1989) Mol. Gen. Genetics 218:78-86), and thus that multiple events must be screened in order to obtain lines displaying the desired expression level and pattern. Such screening may be accomplished by Southern analysis of DNA, Northern analysis of MRNA expression, Western analysis of protein expression, or phenotypic analysis.

For some applications it may be useful to direct the instant sugar transport proteins to different cellular compartments, or to facilitate its secretion from the cell. It is thus envisioned that the chimeric gene described above may be further supplemented by altering the coding sequence to encode Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins with appropriate intracellular targeting sequences such as transit sequences (Keegstra, K. (1989) Cell 56:247-253), signal sequences or sequences encoding endoplasmic reticulum localization (Chrispeels, J. J., (1991) Ann. Rev. Plant Phys. Plant Mol. Biol. 42:21-53), or nuclear localization signals (Raikhel, N. (1992) Plant Phys. 100:1627-1632) added and/or with targeting sequences that are already present removed. While the references cited give examples of each of these, the list is not exhaustive and more targeting signals of utility may be discovered in the future.

It may also be desirable to reduce or eliminate expression of genes encoding Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins in plants for some applications. In order to accomplish this, a chimeric gene designed for co-suppression of the instant sugar transport proteins can be constructed by linking a gene or gene fragment encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein to plant promoter sequences. Alternatively, a chimeric gene designed to express antisense RNA for all or part of the instant nucleic acid fragment can be constructed by linking the gene or gene fragment in reverse orientation to plant promoter sequences. Either the co-suppression or antisense chimeric genes could be introduced into plants via transformation wherein expression of the corresponding endogenous genes are reduced or eliminated.

The instant Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins (or portions thereof) may be produced in heterologous host cells, particularly in the cells of microbial hosts, and can be used to prepare antibodies to the these proteins by methods well known to those skilled in the art. The antibodies are useful for detecting Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins in situ in cells or in vitro in cell extracts. Preferred heterologous host cells for production of the instant sugar transport proteins are microbial hosts. Microbial expression systems and expression vectors containing regulatory sequences that direct high level expression of foreign proteins are well known to those skilled in the art. Any of these could be used to construct a chimeric gene for production of the instant Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins. This chimeric gene could then be introduced into appropriate microorganisms via transformation to provide high level expression of the encoded sugar transport protein. An example of a vector for high level expression of the instant Arabidopsis thaliana-like sugar transport proteins or Beta vulgaris-like sugar transport proteins in a bacterial host is provided (Example 7).

All or a substantial portion of the nucleic acid fragments of the instant invention may also be used as probes for genetically and physically mapping the genes that they are a part of, and as markers for traits linked to those genes. Such information may be useful in plant breeding in order to develop lines with desired phenotypes. For example, the instant nucleic acid fragments may be used as restriction fragment length polymorphism (RFLP) markers. Southern blots (Maniatis) of restriction-digested plant genomic DNA may be probed with the nucleic acid fragments of the instant invention. The resulting banding patterns may then be subjected to genetic analyses using computer programs such as MapMaker (Lander et al., (1987) Genomics 1:174-181) in order to construct a genetic map. In addition, the nucleic acid fragments of the instant invention may be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs of a set of individuals representing parent and progeny of a defined genetic cross. Segregation of the DNA polymorphisms is noted and used to calculate the position of the instant nucleic acid sequence in the genetic map previously obtained using this population (Botstein, D. et al., (1980) Am. J. Hum. Genet. 32:314-331).

The production and use of plant gene-derived probes for use in genetic mapping is described in R. Bernatzky, R. and Tanksley, S. D. (1986) Plant Mol. Biol. Reporter 4(1):37-41. Numerous publications describe genetic mapping of specific cDNA clones using the methodology outlined above or variations thereof. For example, F2 intercross populations, backcross populations, randomly mated populations, near isogenic lines, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes derived from the instant nucleic acid sequences may also be used for physical mapping (i.e., placement of sequences on physical maps; see Hoheisel, J. D., et al., In: Nonmammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and references cited therein).

In another embodiment, nucleic acid probes derived from the instant nucleic acid sequences may be used in direct fluorescence in situ hybridization (FISH) mapping (Trask, B. J. (1991) Trends Genet. 7:149-154). Although current methods of FISH mapping favor use of large clones (several to several hundred KB; see Laan, M. et al. (1995) Genome Research 5:13-20), improvements in sensitivity may allow performance of FISH mapping using shorter probes.

A variety of nucleic acid amplification-based methods of genetic and physical mapping may be carried out using the instant nucleic acid sequences. Examples include allele-specific amplification (Kazazian, H. H. (1989) J. Lab. Clin. Med. 114(2):95-96), polymorphism of PCR-amplified fragments (CAPS; Sheffield, V. C. et al. (1993) Genomics 16:325-332), allele-specific ligation (Landegren, U. et al. (1988) Science 241:1077-1080), nucleotide extension reactions (Sokolov, B. P. (1990) Nucleic Acid Res. 18:3671), Radiation Hybrid Mapping (Walter, M. A. et al. (1997) Nature Genetics 7:22-28) and Happy Mapping (Dear, P. H. and Cook, P. R. (1989) Nucleic Acid Res. 17:6795-6807). For these methods, the sequence of a nucleic acid fragment is used to design and produce primer pairs for use in the amplification reaction or in primer extension reactions. The design of such primers is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify DNA sequence differences between the parents of the mapping cross in the region corresponding to the instant nucleic acid sequence. This, however, is generally not necessary for mapping methods.

Loss of function mutant phenotypes may be identified for the instant cDNA clones either by targeted gene disruption protocols or by identifying specific mutants for these genes contained in a maize population carrying mutations in all possible genes (Ballinger and Benzer, (1989) Proc. Natl. Acad. Sci USA 86:9402; Koes et al., (1995) Proc. Natl. Acad. Sci USA 92:8149; Bensen et al., (1995) Plant Cell 7:75). The latter approach may be accomplished in two ways. First, short segments of the instant nucleic acid fragments may be used in polymerase chain reaction protocols in conjunction with a mutation tag sequence primer on DNAs prepared from a population of plants in which Mutator transposons or some other mutation-causing DNA element has been introduced (see Bensen, supra). The amplification of a specific DNA fragment with these primers indicates the insertion of the mutation tag element in or near the plant gene encoding the Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein. Alternatively, the instant nucleic acid fragment may be used as a hybridization probe against PCR amplification products generated from the mutation population using the mutation tag sequence primer in conjunction with an arbitrary genomic site primer, such as that for a restriction enzyme site-anchored synthetic adaptor. With either method, a plant containing a mutation in the endogenous gene encoding an Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein can be identified and obtained. This mutant plant can then be used to determine or confirm the natural function of the Arabidopsis thaliana-like sugar transport protein or Beta vulgaris-like sugar transport protein gene product.

EXAMPLES

The present invention is further defined in the following Examples, in which all parts and percentages are by weight and degrees are Celsius, unless otherwise stated. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Example 1 Composition of cDNA Libraries: Isolation and Sequencing of cDNA Clones

cDNA libraries representing mRNAs from various corn, rice, soybean and wheat tissues were prepared. The characteristics of the libraries are described below.

TABLE 2 cDNA Libraries from Corn, Rice, Soybean and Wheat Library Tissue Clone cc1 Corn (Zea mays L.) callus stage 1** cc1.mn0002.h1 Cepe7 Corn (Zea mays L.) epicotyl from 7 day old etiolated cepe7.pk0018.g3 seedling cil1c Corn (Zea mays L.) pooled immature leaf tissue at V4, cil1c.pk001.f21 V6 and V8** cr1n Corn (Zea mays L.) root from 7 day seedlings grown in cr1n.pk0143.h10 light* p0002 Corn (Zea mays L.) tassel: premeiotic > early uninucleate p0002.cgevb73r p0032 Corn (Zea mays L.) regenernerating callus, 10 and 14 days p0032.crcba66r after auxin removal. p0072 Corn (Zea mays L.) 14 days after planting etiolated p0072.comgi92r seedling: mesocotyl p0097 Corn (Zea mays L.) V9, 7 cm whorl section after p0097.cqran4lr application of European Corn Borer p0106 Corn (Zea mays L.) 5 days after pollenation whole kernels* p0106.cjlpm67r p0114 Corn (Zea mays L.) intercalary meristem of expanding p0114.cimm181r internodes 5-9 at V10 stage* p0128 Corn (Zea mays L.) pooled primary and secondary p0128.cpict38 immature ear Rdslc Rice (Oryza sativa, YM) developing seeds rdslc.pk007.n17 rlr6 Rice (Oryza sativa L.) leaf (15 days after germination) rlr6.pk0005.b10 6 hrs after infection of Magaporthe grisea strain 4360-R-62 (AVR2-YAMO); Resistant r10n Rice (Oryza sativa L.) 15 day leaf* r10n.pk102.p24 r10n.pk107.p2 rlr12 Rice (Oryza sativa L.) leaf, 15 days after germination, rlr12.pk0013.d11 12 hours after infection of Magaporthe grisea strain 4360-R-62 (AVR2-YAMO); Resistant rls6 Rice (Oryza sativa L.) leaf, 15 days after germination, rls6.pk0003.d5 6 hrs after infection of Magaporthe grisea strain 4360-R-67 (avr2-yamo); Susceptible sdp3c Soybean (Glycine max L.) developing pods 8-9 mm sdp3c.pk012.i1 sfl1 Soybean (Glycine max L.) immature flower sfl1.pk0079.a4 sfl1.pk0058.h12 sgs2c Soybean (Glycine max L.) seeds 14 hrs after germination sgs2c.pk004.o17 sgs4c Soybean (Glycine max L.) seeds 2 days after germination sgs4c.pk005.c9 srl Soybean (Glycine max L.) root library srl.pk0061.g8 sre Soybean (Glycine max L.) root elongation sre.pk0032.h6 ssl Soybean (Glycine max L.) seedling 5-10 day ssl.pk0022.f1 wlk8 Wheat (Triticum aestivum L.) seedlings 8 hr after wlk8.pk0001.a11 treatment with fungicide*** wlk8.pk0001.a12 wlm1 Wheat (Triticum aestivum L.) seedlings 1 hr after wlm1.pk0012.h1 inoculation with Erysiphe graminis f. sp tritici wlm96 Wheat (Triticum aestivum L.) seedlings 96 hr after wlm96.pk043.e19 inoculation w/ E. graminis wre1n Wheat (Triticum aestivum L.) root; 7 day old etiolated wre1n.pk0006.b4 seedling* wre1n.pk0062.g6 *These libraries were normalized essentially as described in U.S. Pat. No. 5,482,845 **V4, V6 and V8 refer to stages of corn growth. The descriptions can be found in “How a Corn Plant Develops” Special Report No. 48, Iowa State University of Science and Technology Cooperative Extension Service Ames, Iowa, Reprinted February 1996. ***Application of 6-iodo-2-propoxy-3-propyl-4(3H-quinazolinone; synthesis and methods of using this compound are described in USSN 08/545,827, incorporated herein by reference.

cDNA libraries were prepared in Uni-ZAP™ XR vectors according to the manufacturer's protocol (Stratagene Cloning Systems, La Jolla, Calif.). Conversion of the Uni-ZAP™ XR libraries into plasmid libraries was accomplished according to the protocol provided by Stratagene. Upon conversion, cDNA inserts were contained in the plasmid vector pBluescript. cDNA inserts from randomly picked bacterial colonies containing recombinant pBluescript plasmids were amplified via polymerase chain reaction using primers specific for vector sequences flanking the inserted cDNA sequences or plasmid DNA was prepared from cultured bacterial cells. Amplified insert DNAs or plasmid DNAs were sequenced in dye-primer sequencing reactions to generate partial cDNA sequences (expressed sequence tags or “ESTs”; see Adams, M. D. et al., (1991) Science 252:1651). The resulting ESTs were analyzed using a Perkin Elmer Model 377 fluorescent sequencer.

Example 2 Identification of cDNA Clones

ESTs encoding sugar transport proteins were identified by conducting BLAST (Basic Local Alignment Search Tool; Altschul, S. F., et al., (1993) J. Mol. Biol. 215:403-410; see also www.ncbi.nlm.nih.gov/BLAST/) searches for similarity to sequences contained in the BLAST “nr” database (comprising all non-redundant GenBank CDS translations, sequences derived from the 3-dimensional structure Brookhaven Protein Data Bank, the last major release of the SWISS-PROT protein sequence database, EMBL, and DDBJ databases). The cDNA sequences obtained in Example 1 were analyzed for similarity to all publicly available DNA sequences contained in the “nr” database using the BLASTN algorithm provided by the National Center for Biotechnology Information (NCBI). The DNA sequences were translated in all reading frames and compared for similarity to all publicly available protein sequences contained in the “nr” database using the BLASTX algorithm (Gish, W. and States, D. J. (1993) Nature Genetics 3:266-272 and Altschul, Stephen F., et al. (1997) Nucleic Acids Res. 25:3389-3402) provided by the NCBI. For convenience, the P-value (probability) of observing a match of a cDNA sequence to a sequence contained in the searched databases merely by chance as calculated by BLAST are reported herein as “pLog” values, which represent the negative of the logarithm of the reported P-value. Accordingly, the greater the pLog value, the greater the likelihood that the cDNA sequence and the BLAST “hit” represent homologous proteins.

Example 3 Characterization of cDNA Clones Encoding Arabidopsis thaliana-Like Sugar Transport Proteins

The BLASTX search using the EST sequences from several corn, rice, soybean and wheat clones revealed similarity of the proteins encoded by the cDNAs to a sugar transport protein from Arabidopsis thaliana (NCBI Identifier No. gi 3080420). In the process of comparing the ESTs it was found that many of the clones had overlapping regions of homology. Using this homology it was possible to align the ESTs and assemble several contigs encoding unique corn, rice, soybean and wheat sugar transport proteins. The individual clones and the composition of each assembled contig are shown in Table 3. The BLAST results for each of the contigs and individual ESTs and are also shown in Table 3:

TABLE 3 BLAST Results for Clones Encoding Polypeptides Homologous to Arabidopsis thaliana Sugar Transport Protein Clone BLAST pLog Score Contig composed of clones: >250.00 p0032.crcba66r p0097.cqran41r crln.pk0143.h10 p0128.cpict38 p0106.cjlpm67r cillc.pk001.f21 p0072.comgi92r p0114.cimm181r p0002.cgcvb73r Contig composed of clones: 27.70 rlr12.pk0013.d11 rdslc.pk007.n17 rls6.pk0003.d5 54.00 Contig composed of clones: >250.00 sgs4c.pk005.c9 sfl1.pk0079.a4 sdp3c.pk012.i1 ssl.pk0022.fl >250.00 wlk8.pk0001.a12 21.30 Contig composed of clones: 149.00 wlm96.pk043.el9 wreln.pk0062.g6 wreln.pk0006.b4 117.00

The sequence of the corn contig composed of clones p0032.crcba66r, p0097.cqran41r, cr1n.pk0143.h10, p0128.cpict38, p0106.cjlpm67r, cil1c.pk001.f21, p0072.comgi92p0114 .cimm181r and p0002.cgevb73r is shown in SEQ ID NO:1; the deduced amino acid sequence of this contig, which represents 100% of the protein, is shown in SEQ ID NO:2. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:2 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:2 is 66% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of the rice contig composed of clones rlr12.pk0013.d11 and rds1c.pk007.n17 is shown in SEQ ID NO:3; the deduced amino acid sequence of this contig, which represents 9% of the protein (N-terminal region), is shown in SEQ ID NO:4. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:4 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:2 is 86% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of the entire cDNA insert from clone rls6.pk0003.d5 is shown in SEQ ID NO:5; the deduced amino acid sequence of this cDNA, which represents 18% of the of the protein (C-terminal region), is shown in SEQ ID NO:6. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:6 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:6 is 74% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of the soybean contig composed of clones sgs4c.pk005.c9, sfl1.pk0079.a4 and sdp3c.pk012.i1 is shown in SEQ ID NO:7; the deduced amino acid sequence of this contig, which represents 100% of the protein, is shown in SEQ ID NO:8. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:8 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:8 is 68% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of a portion of the cDNA insert from clone ss1.pk0022.f1 is shown in SEQ ID NO:9; the deduced amino acid sequence of this cDNA, which represents 66% of the of the protein (C-terminal region), is shown in SEQ ID NO:10. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:10 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:10 is 66% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of a portion of the cDNA insert from clone wlk8.pk0001.a12 is shown in SEQ ID NO:11; the deduced amino acid sequence of this cDNA, which represents 7% of the of the protein (N-terminal region), is shown in SEQ ID NO:12. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:12 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:12 is 88% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of the wheat contig composed of clones wlm96.pk043.e19 and wre1n.pk0062.g6 is shown in SEQ ID NO:13; the deduced amino acid sequence of this contig, which represents 45% of the protein (C-terminal region), is shown in SEQ ID NO:14. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:14 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:14 is 65% similar to the Arabidopsis thaliana sugar transport protein.

The sequence of a portion of the cDNA insert from clone wre1n.pk0006.b4 is shown in SEQ ID NO:15; the deduced amino acid sequence of this cDNA, which represents 31% of the of the protein (C-terminal region), is shown in SEQ ID NO:16. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:16 and the Arabidopsis thaliana sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:16 is 76% similar to the Arabidopsis thaliana sugar transport protein.

FIG. 1 presents an alignment of the amino acid sequence set forth in SEQ ID NOS:2, 4, 6, 8, 10, 12, 14 and 16 with the Arabidopsis thaliana-like sugar transport protein amino acid sequence, SEQ ID NO:29. Alignments were performed using the Clustal algorithm. The percent similarity between the corn, rice, soybean and wheat acid sequences was calculated to range between 16% to 89% using the Clustal algorithm.

BLAST scores and probabilities indicate that the instant nucleic acid fragments encode portions of sugar transport proteins. These sequences represent the first corn, rice, soybean and wheat sequences encoding Arabidopsis thaliana-like sugar transport proteins.

Example 4 Characterization of cDNA Clones Encoding Beta vulgaris-Like Sugar Transport Proteins

The BLASTX search using the EST sequences from several corn, rice, soybean and wheat clones revealed similarity of the proteins encoded by the cDNAs to a sugar transport protein from Beta vulgaris (NCBI Identifier No. gi 1778093). In the process of comparing the ESTs it was found that several of the rice and soybean clones had overlapping regions of homology. Using this homology it was possible to align the ESTs and assemble two contigs encoding unique rice and soybean B. vulgaris-like sugar transport proteins. The individual clones and the assembled composition of each contig are shown in Table 4. The BLAST results for each of the contigs and individual ESTs and are also shown in Table 4:

TABLE 4 BLAST Results for Clones Encoding Polypeptides Homologous to Beta vulgaris Sugar Transport Protein Clone BLAST pLog Score ccl.mn0002.h1 53.70 cepe7.pk0018.g3 164.00 Contig composed of clones: >250.00 rlr6.pk0005.b10 rl0n.pk102.p24 rl0n.pk107.p2 Contig composed of clones: >250.00 srl.pk0061.g8 sfl1.pk0058.h12 sgs2c.pk004.o17 sre.pk0032.h6 wlk8.pk0001.a11 >250.00 wlml.pk0012.h1 >250.00

The sequence of a portion of the cDNA insert from clone cc1.mn0002.h1 is shown in SEQ ID NO:17; the deduced amino acid sequence of this cDNA, which represents 31% of the of the protein (N-terminal region), is shown in SEQ ID NO:18. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:18 and the Beta vulgaris sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:18 is 65% similar to the Beta vulgaris sugar transport protein.

The sequence of the entire cDNA insert from clone cepe7.pk0018.g3 is shown in SEQ ID NO:19; the deduced amino acid sequence of this cDNA, which represents 100% of the of the protein, is shown in SEQ ID NO:20. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:20 and the Beta vulgaris sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:20 is 57% similar to the Beta vulgaris sugar transport protein.

The sequence of the rice contig composed of clones rlr6.pk0005.b10, r10n.pk102.p24 and r10n.pk107.p2 is shown in SEQ ID NO:21; the deduced amino acid sequence of this contig, which represents 100% of the protein, is shown in SEQ ID NO:22. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:22 and the Beta vulgaris sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:22 is 61% similar to the Beta vulgaris sugar transport protein.

The sequence of the soybean contig composed of clones sr1.pk0061.g8, sfl1.pk0058.h12, sgs2c.pk004.o17 and sre.pk0032.h6 is shown in SEQ ID NO:23; the deduced amino acid sequence of this contig, which represents 100% of the protein, is shown in SEQ ID NO :24. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:24 and the Beta vulgaris sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:24 is 66% similar to the Beta vulgaris sugar transport protein.

The sequence of the entire cDNA insert from clone wlk8.pk0001.a11 is shown in SEQ ID NO:25; the deduced amino acid sequence of this cDNA, which represents 100% of the of the protein, is shown in SEQ ID NO:26. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:26 and the Beta vulgaris sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:26 is 61% similar to the Beta vulgaris sugar transport protein.

The sequence of the entire cDNA insert from clone wlm1.pk0012.h1 is shown in SEQ ID NO:27; the deduced amino acid sequence of this cDNA, which represents 100% of the of the protein, is shown in SEQ ID NO:28. A calculation of the percent similarity of the amino acid sequence set forth in SEQ ID NO:28 and the Beta vulgaris sequence (using the Clustal algorithm) revealed that the protein encoded by SEQ ID NO:28 is 56% similar to the Beta vulgaris sugar transport protein.

FIG. 2 presents an alignment of the amino acid sequence set forth in SEQ ID NOS:18, 20, 22, 24, 26 and 28 with the Beta vulgaris-like sugar transport protein amino acid sequence, SEQ ID NO:30. Alignments were performed using the Clustal algorithm. The percent similarity between the corn, rice, soybean and wheat acid sequences was calculated to range between 43% to 81% using the Clustal algorithm.

BLAST scores and probabilities indicate that the instant nucleic acid fragments encode portions of sugar transport proteins. These sequences represent the first corn, rice, soybean and wheat sequences encoding Beta vulgaris-like sugar transport proteins.

Example 5 Expression of Chimeric Genes in Monocot Cells

A chimeric gene comprising a cDNA encoding sugar transport protein in sense orientation with respect to the maize 27 kD zein promoter that is located 5′ to the cDNA fragment, and the 10 kD zein 3′ end that is located 3′ to the cDNA fragment, can be constructed. The cDNA fragment of this gene may be generated by polymerase chain reaction (PCR) of the cDNA clone using appropriate oligonucleotide primers. Cloning sites (NcoI or SmaI) can be incorporated into the oligonucleotides to provide proper orientation of the DNA fragment when inserted into the digested vector pML 103 as described below. Amplification is then performed in a standard PCR. The amplified DNA is then digested with restriction enzymes NcoI and SmaI and fractionated on an agarose gel. The appropriate band can be isolated from the gel and combined with a 4.9 kb NcoI-SmaI fragment of the plasmid pML103. Plasmid pML103 has been deposited under the terms of the Budapest Treaty at ATCC (American Type Culture Collection, 10801 University Blvd., Manassas, Va. 20110-2209), and bears accession number ATCC 97366. The DNA segment from pML103 contains a 1.05 kb SalI-NcoI promoter fragment of the maize 27 kD zein gene and a 0.96 kb SmaI-SalI fragment from the 3′ end of the maize 10 kD zein gene in the vector pGem9Zf(+) (Promega). Vector and insert DNA can be ligated at 15° C. overnight, essentially as described (Maniatis). The ligated DNA may then be used to transform E. coli XL1-Blue (Epicurian Coli XL-1 Blue™M; Stratagene). Bacterial transformants can be screened by restriction enzyme digestion of plasmid DNA and limited nucleotide sequence analysis using the dideoxy chain termination method (Sequenase™ DNA Sequencing Kit; U.S. Biochemical). The resulting plasmid construct would comprise a chimeric gene encoding, in the 5′ to 3′ direction, the maize 27 kD zein promoter, a cDNA fragment encoding a sugar transport protein, and the 10 kD zein 3′ region.

The chimeric gene described above can then be introduced into corn cells by the following procedure. Immature corn embryos can be dissected from developing caryopses derived from crosses of the inbred corn lines H99 and LH132. The embryos are isolated 10 to 11 days after pollination when they are 1.0 to 1.5 mm long. The embryos are then placed with the axis-side facing down and in contact with agarose-solidified N6 medium (Chu et al., (1975) Sci. Sin. Peking 18:659-668). The embryos are kept in the dark at 27° C. Friable embryogenic callus consisting of undifferentiated masses of cells with somatic proembryoids and embryoids borne on suspensor structures proliferates from the scutellum of these immature embryos. The embryogenic callus isolated from the primary explant can be cultured on N6 medium and sub-cultured on this medium every 2 to 3 weeks.

The plasmid, p35S/Ac (obtained from Dr. Peter Eckes, Hoechst Ag, Frankfurt, Germany) may be used in transformation experiments in order to provide for a selectable marker. This plasmid contains the Pat gene (see European Patent Publication 0 242 236) which encodes phosphinothricin acetyl transferase (PAT). The enzyme PAT confers resistance to herbicidal glutamine synthetase inhibitors such as phosphinothricin. The pat gene in p35S/Ac is under the control of the 35S promoter from Cauliflower Mosaic Virus (Odell et al. (1985) Nature 313:810-812) and the 3′ region of the nopaline synthase gene from the T-DNA of the Ti plasmid of Agrobacterium tumefaciens.

The particle bombardment method (Klein et al., (1987) Nature 327:70-73) may be used to transfer genes to the callus culture cells. According to this method, gold particles (1 μm in diameter) are coated with DNA using the following technique. Ten μg of plasmid DNAs are added to 50 μL of a suspension of gold particles (60 mg per mL). Calcium chloride (50 μL of a 2.5M solution) and spermidine free base (20 μL of a 1.0M solution) are added to the particles. The suspension is vortexed during the addition of these solutions. After 10 minutes, the tubes are briefly centrifuged (5 sec at 15,000 rpm) and the supernatant removed. The particles are resuspended in 200 μL of absolute ethanol, centrifuged again and the supernatant removed. The ethanol rinse is performed again and the particles resuspended in a final volume of 30 μL of ethanol. An aliquot (5 μL) of the DNA-coated gold particles can be placed in the center of a Kaptonh™ flying disc (Bio-Rad Labs). The particles are then accelerated into the corn tissue with a Biolistic™ PDS-1000/He (Bio-Rad Instruments, Hercules Calif.), using a helium pressure of 1000 psi, a gap distance of 0.5 cm and a flying distance of 1.0 cm.

For bombardment, the embryogenic tissue is placed on filter paper over agarose-solidified N6 medium. The tissue is arranged as a thin lawn and covered a circular area of about 5 cm in diameter. The petri dish containing the tissue can be placed in the chamber of the PDS-1000/He approximately 8 cm from the stopping screen. The air in the chamber is then evacuated to a vacuum of 28 inches of Hg. The macrocarrier is accelerated with a helium shock wave using a rupture membrane that bursts when the He pressure in the shock tube reaches 1000 psi.

Seven days after bombardment the tissue can be transferred to N6 medium that contains gluphosinate (2 mg per liter) and lacks casein or proline. The tissue continues to grow slowly on this medium. After an additional 2 weeks the tissue can be transferred to fresh N6 medium containing gluphosinate. After 6 weeks, areas of about 1 cm in diameter of actively growing callus can be identified on some of the plates containing the glufosinate-supplemented medium. These calli may continue to grow when sub-cultured on the selective medium.

Plants can be regenerated from the transgenic callus by first transferring clusters of issue to N6 medium supplemented with 0.2 mg per liter of 2,4-D. After two weeks the issue can be transferred to regeneration medium (Fromm et al., (1990) Bio/Technology 8:833-839).

Example 6 Expression of Chimeric Genes in Dicot Cells

A seed-specific expression cassette composed of the promoter and transcription terminator from the gene encoding the β subunit of the seed storage protein phaseolin from the bean Phaseolus vulgaris (Doyle et al. (1986) J. Biol. Chem. 261:9228-9238) can be used for expression of the instant sugar transport proteins in transformed soybean. The phaseolin cassette includes about 500 nucleotides upstream (5′ ) from the translation initiation codon and about 1650 nucleotides downstream (3′ ) from the translation stop codon of phaseolin. Between the 5′ and 3′ regions are the unique restriction endonuclease sites Nco I (which includes the ATG translation initiation codon), Sma I, Kpn I and Xba I. The entire cassette is flanked by Hind III sites.

The cDNA fragment of this gene may be generated by polymerase chain reaction (PCR) of the cDNA clone using appropriate oligonucleotide primers. Cloning sites can be incorporated into the oligonucleotides to provide proper orientation of the DNA fragment when inserted into the expression vector. Amplification is then performed as described above, and the isolated fragment is inserted into a pUC18 vector carrying the seed expression cassette.

Soybean embroys may then be transformed with the expression vector comprising a sequence encoding a sugar transport protein. To induce somatic embryos, cotyledons, 3-5 mm in length dissected from surface sterilized, immature seeds of the soybean cultivar A2872, can be cultured in the light or dark at 26° C. on an appropriate agar medium for 6-10 weeks. Somatic embryos which produce secondary embryos are then excised and placed into a suitable liquid medium. After repeated selection for clusters of somatic embryos which multiplied as early, globular staged embryos, the suspensions are maintained as described below.

Soybean embryogenic suspension cultures can maintained in 35 mL liquid media on a rotary shaker, 150 rpm, at 26° C. with florescent lights on a 16:8 hour day/night schedule. Cultures are subcultured every two weeks by inoculating approximately 35 mg of tissue into 35 mL of liquid medium.

Soybean embryogenic suspension cultures may then be transformed by the method of particle gun bombardment (Kline et al. (1987) Nature (London) 327:70, U.S. Pat. No. 4,945,050). A DuPont Biolistic™ PDS1000/HE instrument (helium retrofit) can be used for these transformations.

A selectable marker gene which can be used to facilitate soybean transformation is a chimeric gene composed of the 35S promoter from Cauliflower Mosaic Virus (Odell et al. (1985) Nature 313:810-812), the hygromycin phosphotransferase gene from plasmid pJR225 (from E. coli; Gritz et al. (1983) Gene 25:179-188) and the 3′ region of the nopaline synthase gene from the T-DNA of the Ti plasmid of Agrobacterium tumefaciens. The seed expression cassette comprising the phaseolin 5′ region, the fragment encoding the sugar transport protein and the phaseolin 3′ region can be isolated as a restriction fragment. This fragment can then be inserted into a unique restriction site of the vector carrying the marker gene.

To 50 μL of a 60 mg/mL 1 μm gold particle suspension is added (in order): 5 μL DNA (1 μg/μL), 20 μl spermidine (0.1 M), and 50 μL CaCl₂ (2.5 M). The particle preparation is then agitated for three minutes, spun in a microfage for 10 seconds and the supernatant removed. The DNA-coated particles are then washed once in 400 μL 70% ethanol and resuspended in 40 μL of anhydrous ethanol. The DNA/particle suspension can be sonicated three times for one second each. Five μL of the DNA-coated gold particles are then loaded on each macro carrier disk.

Approximately 300-400 mg of a two-week-old suspension culture is placed in an empty 60×15 mm petri dish and the residual liquid removed from the tissue with a pipette. For each transformation experiment, approximately 5-10 plates of tissue are normally bombarded. Membrane rupture pressure is set at 1100 psi and the chamber is evacuated to a vacuum of 28 inches mercury. The tissue is placed approximately 3.5 inches away from the retaining screen and bombarded three times. Following bombardment, the tissue can be divided in half and placed back into liquid and cultured as described above.

Five to seven days post bombardment, the liquid media may be exchanged with fresh media, and eleven to twelve days post bombardment with fresh media containing 50 mg/mL hygromycin. This selective media can be refreshed weekly. Seven to eight weeks post bombardment, green, transformed tissue may be observed growing from untransformed, necrotic embryogenic clusters. Isolated green tissue is removed and inoculated into individual flasks to generate new, clonally propagated, transformed embryogenic suspension cultures. Each new line may be treated as an independent transformation event. These suspensions can then be subcultured and maintained as clusters of immature embryos or regenerated into whole plants by maturation and germination of individual somatic embryos.

Example 7 Expression of Chimeric Genes in Microbial Cells

The cDNAs encoding the instant sugar transport proteins can be inserted into the T7 E. coli expression vector pBT430. This vector is a derivative of pET-3a (Rosenberg et al. (1987) Gene 56:125-135) which employs the bacteriophage T7 RNA polymerase/T7 promoter system. Plasmid pBT430 was constructed by first destroying the EcoR I and Hind III sites in pET-3 a at their original positions. An oligonucleotide adaptor containing EcoR I and Hind III sites was inserted at the BamH I site of pET-3a. This created pET-3aM with additional unique cloning sites for insertion of genes into the expression vector. Then, the Nde I site at the position of translation initiation was converted to an Nco I site using ligonucleotide-directed mutagenesis. The DNA sequence of pET-3aM in this region, 5′-CATATGG, was converted to 5′-CCCATGG in pBT430.

Plasmid DNA containing a cDNA may be appropriately digested to release a nucleic acid fragment encoding the protein. This fragment may then be purified on a 1% NuSieve GTGTM low melting agarose gel (FMC). Buffer and agarose contain 10 μg/ml ethidium bromide for visualization of the DNA fragment. The fragment can then be purified from the agarose gel by digestion with GELase™ (Epicentre Technologies) according to the manufacturer's instructions, ethanol precipitated, dried and resuspended in 20 μL of water. Appropriate oligonucleotide adapters may be ligated to the fragment using T4 DNA ligase (New England Biolabs, Beverly, Mass.). The fragment containing the ligated adapters can be purified from the excess adapters using low melting agarose as described above. The vector pBT430 is digested, dephosphorylated with alkaline phosphatase (NEB) and deproteinized with phenol/chloroform as described above. The prepared vector pBT430 and fragment can then be ligated at 16° C. for 15 hours followed by transformation into DH5 electrocompetent cells (GIBCO BRL). Transformants can be selected on agar plates containing LB media and 100 μg/mL ampicillin. Transformants containing the gene encoding the sugar transport protein are then screened for the correct orientation with respect to the T7 promoter by restriction enzyme analysis.

For high level expression, a plasmid clone with the cDNA insert in the correct orientation relative to the T7 promoter can be transformed into E. coli strain BL21(DE3) (Studier et al. (1986) J. Mol. Biol. 189:113-130). Cultures are grown in LB medium containing ampicillin (100 mg/L) at 25° C. At an optical density at 600 nm of approximately 1, IPTG (isopropylthio-β-galactoside, the inducer) can be added to a final concentration of 0.4 mM and incubation can be continued for 3 h at 25°. Cells are then harvested by centrifugation and re-suspended in 50 μL of 50 mM Tris-HCl at pH 8.0 containing 0.1 mM DTT and 0.2 mM phenyl methylsulfonyl fluoride. A small amount of 1 mm glass beads can be added and the mixture sonicated 3 times for about 5 seconds each time with a microprobe sonicator. The mixture is centrifuged and the protein concentration of the supernatant determined. One μg of protein from the soluble fraction of the culture can be separated by SDS-polyacrylamide gel electrophoresis. Gels can be observed for protein bands migrating at the expected molecular weight.

30 1 2824 DNA Zea mays unsure (29) unsure (622) unsure (636) unsure (638) unsure (669) unsure (771) unsure (822) unsure (856) unsure (889) unsure (896) unsure (944) 1 cccacccccc tccactccac taccacggng gcacggcctg cctctgcagc tctgccctgc 60 tccgcacccc tcgctctcca accccaacgc gcggcgttgc taaaattcac ctcagcgcgt 120 actccagttt ggccacctca ccacccgccg ccgctgttta agaaggcccc gcgcccgatc 180 ggggatcacg aaccttggcc gccgctgccg gagtgggggc gtagatttcc ggcggccatg 240 gggggcgccg tgatggtcgc catcgcggcc tctatcggca acttgctgca gggctgggac 300 aatgcgacaa ttgctggagc cgtcctgtac ataaagaagg aattcaacct gcagagcgag 360 cctctgatcg aaggcctcat cgtcgccatg ttcctcattg gggcaacagt catcacaaca 420 tctccggggc caagggctga ctgcgttggt aggaggccca tgctggtcgc ctcggctgtc 480 ctctacttcg tcagtgggct ggtgatgctt tgggcgccaa ttgtgtacat cttgctcctc 540 gcaaggctca ttgatgggtt cggtatcggt ttggcggtca cacttgttcc tctctacatc 600 tccgaaactg caccgcacag anattcttgg ggctgntnga acacgttgcc gcagttcatt 660 ggggtcagng gagggatgtt cctctcctac tgcatggtgt ttgggatgtc cctcatgccc 720 aaacctgatt ggaggctcat gcttggagtt ctgtcgatcc cgtcacttat ntactttgga 780 ctgactgtct tctacttgcc tgaatcacca aggtggcttg tnagcaaagg aaggatggcg 840 gaggcgaaga gagtgntgca aaggctgcgg ggaagagaag atgtctcang ggaganggct 900 cttctagttg aaggtttggg ggtcggtaaa gatacacgta tttnagagta catcattgga 960 cctgccaccg aggcagccga tgatcttgta actgacggtg ataaggaaca aatcacactt 1020 tatgggcctg aagaaggcca gtcatggatt gctcgacctt ctaagggacc catcatgctt 1080 ggaagtgtgc tttctcttgc atctcgtcat gggagcatgg tgaaccagag tgtacccctt 1140 atggatccga ttgtgacact ttttggtagt gtccatgaga atatgcctca agctggagga 1200 agtatgagga gcacattgtt tccaaacttt ggaagtatgt tcagtgtcac agatcagcat 1260 gccaaaaatg agcagtggga tgaagagaat cttcataggg atgacgagga gtacgcatct 1320 gatggtgcag gaggtgacta tgaggacaat ctccatagcc cattgctgtc caggcaggca 1380 acaggtgcgg aagggaagga cattgtgcac catggtcacc gtggaagtgc tttgagcatg 1440 agaaggcaaa gcctcttagg ggagggtgga gatggtgtga gcagcactga tatcggtggg 1500 ggatggcagc ttgcttggaa atggtcagag aaggaaggtg agaatggtag aaaggaaggt 1560 ggtttcaaaa gagtctactt gcaccaagag ggagttcctg gctcaagaag gggctcaatt 1620 gtttcacttc ccggtggtgg cgatgttctt gagggtagtg agtttgtaca tgctgctgct 1680 ttagtaagtc agtcagcact tttctcaaag ggtcttgctg aaccacgcat gtcagatgct 1740 gccatggttc acccatctga ggtagctgcc aaaggttcac gttggaaaga tttgtttgaa 1800 cctggagtga ggcgtgccct gttagtcggt gttggaattc agatccttca acagtttgct 1860 ggaataaacg gtgttctgta ctatacccca caaattcttg agcaagctgg tgtggcagtt 1920 attctttcca aatttggtct cagctcggca tcagcatcca tcttgatcag ttctctcact 1980 accttactaa tgcttccttg cattggcttt gccatgctgc ttatggatct ttccggaaga 2040 aggtttttgc tgctaggcac aattccaatc ttgatagcat ctctagttat cctggttgtg 2100 tccaatctaa ttgatttggg tacactagcc catgctttgc tctccaccat cagtgttatc 2160 gtctacttct gctgcttcgt tatgggattt ggtcccatcc ccaacatttt atgtgcagag 2220 atctttccaa ccagggttcg tggcctctgt attgccattt gtgcctttac attctggatc 2280 ggagatatca tcgtcaccta cagccttcct gtgatgctga atgctattgg actggcgggt 2340 gttttcagca tatatgcagt cgtatgcttg atttcctttg tgttcgtctt ccttaaggtc 2400 cctgagacaa aggggatgcc ccttgaggtt attaccgaat tctttgcagt tggtgcgaag 2460 caagcggctg caaaagccta atttctttgg tacctttgtg tgcaactatt gcactgtaag 2520 ttagaaactt gaaggggttt caccaagaag ctcggagaat tactttggat ttgtgtaaat 2580 gttaagggaa cgaacatctg ctcatgctcc tcaaacggta aaaaagagtc cctcaatggc 2640 aaataggagt cgttaagttg tcaatgtcat ttaccatatg ttttacctat ttgtactgta 2700 ttataagtca agctattcaa cgctggttgt tgctagaaat ctttagaaca aagatgataa 2760 tgatctgatc tgatgttata atattcaaat ctcaaataaa gaaaatatcg tttctcaaaa 2820 aaaa 2824 2 747 PRT Zea mays UNSURE (129) UNSURE (133)..(134) UNSURE (144) UNSURE (178) UNSURE (207) UNSURE (218) UNSURE (220) UNSURE (236) 2 Met Gly Gly Ala Val Met Val Ala Ile Ala Ala Ser Ile Gly Asn Leu 1 5 10 15 Leu Gln Gly Trp Asp Asn Ala Thr Ile Ala Gly Ala Val Leu Tyr Ile 20 25 30 Lys Lys Glu Phe Asn Leu Gln Ser Glu Pro Leu Ile Glu Gly Leu Ile 35 40 45 Val Ala Met Phe Leu Ile Gly Ala Thr Val Ile Thr Thr Ser Pro Gly 50 55 60 Pro Arg Ala Asp Cys Val Gly Arg Arg Pro Met Leu Val Ala Ser Ala 65 70 75 80 Val Leu Tyr Phe Val Ser Gly Leu Val Met Leu Trp Ala Pro Ile Val 85 90 95 Tyr Ile Leu Leu Leu Ala Arg Leu Ile Asp Gly Phe Gly Ile Gly Leu 100 105 110 Ala Val Thr Leu Val Pro Leu Tyr Ile Ser Glu Thr Ala Pro His Arg 115 120 125 Xaa Ser Trp Gly Xaa Xaa Asn Thr Leu Pro Gln Phe Ile Gly Val Xaa 130 135 140 Gly Gly Met Phe Leu Ser Tyr Cys Met Val Phe Gly Met Ser Leu Met 145 150 155 160 Pro Lys Pro Asp Trp Arg Leu Met Leu Gly Val Leu Ser Ile Pro Ser 165 170 175 Leu Xaa Tyr Phe Gly Leu Thr Val Phe Tyr Leu Pro Glu Ser Pro Arg 180 185 190 Trp Leu Val Ser Lys Gly Arg Met Ala Glu Ala Lys Arg Val Xaa Gln 195 200 205 Arg Leu Arg Gly Arg Glu Asp Val Ser Xaa Glu Xaa Ala Leu Leu Val 210 215 220 Glu Gly Leu Gly Val Gly Lys Asp Thr Arg Ile Xaa Glu Tyr Ile Ile 225 230 235 240 Gly Pro Ala Thr Glu Ala Ala Asp Asp Leu Val Thr Asp Gly Asp Lys 245 250 255 Glu Gln Ile Thr Leu Tyr Gly Pro Glu Glu Gly Gln Ser Trp Ile Ala 260 265 270 Arg Pro Ser Lys Gly Pro Ile Met Leu Gly Ser Val Leu Ser Leu Ala 275 280 285 Ser Arg His Gly Ser Met Val Asn Gln Ser Val Pro Leu Met Asp Pro 290 295 300 Ile Val Thr Leu Phe Gly Ser Val His Glu Asn Met Pro Gln Ala Gly 305 310 315 320 Gly Ser Met Arg Ser Thr Leu Phe Pro Asn Phe Gly Ser Met Phe Ser 325 330 335 Val Thr Asp Gln His Ala Lys Asn Glu Gln Trp Asp Glu Glu Asn Leu 340 345 350 His Arg Asp Asp Glu Glu Tyr Ala Ser Asp Gly Ala Gly Gly Asp Tyr 355 360 365 Glu Asp Asn Leu His Ser Pro Leu Leu Ser Arg Gln Ala Thr Gly Ala 370 375 380 Glu Gly Lys Asp Ile Val His His Gly His Arg Gly Ser Ala Leu Ser 385 390 395 400 Met Arg Arg Gln Ser Leu Leu Gly Glu Gly Gly Asp Gly Val Ser Ser 405 410 415 Thr Asp Ile Gly Gly Gly Trp Gln Leu Ala Trp Lys Trp Ser Glu Lys 420 425 430 Glu Gly Glu Asn Gly Arg Lys Glu Gly Gly Phe Lys Arg Val Tyr Leu 435 440 445 His Gln Glu Gly Val Pro Gly Ser Arg Arg Gly Ser Ile Val Ser Leu 450 455 460 Pro Gly Gly Gly Asp Val Leu Glu Gly Ser Glu Phe Val His Ala Ala 465 470 475 480 Ala Leu Val Ser Gln Ser Ala Leu Phe Ser Lys Gly Leu Ala Glu Pro 485 490 495 Arg Met Ser Asp Ala Ala Met Val His Pro Ser Glu Val Ala Ala Lys 500 505 510 Gly Ser Arg Trp Lys Asp Leu Phe Glu Pro Gly Val Arg Arg Ala Leu 515 520 525 Leu Val Gly Val Gly Ile Gln Ile Leu Gln Gln Phe Ala Gly Ile Asn 530 535 540 Gly Val Leu Tyr Tyr Thr Pro Gln Ile Leu Glu Gln Ala Gly Val Ala 545 550 555 560 Val Ile Leu Ser Lys Phe Gly Leu Ser Ser Ala Ser Ala Ser Ile Leu 565 570 575 Ile Ser Ser Leu Thr Thr Leu Leu Met Leu Pro Cys Ile Gly Phe Ala 580 585 590 Met Leu Leu Met Asp Leu Ser Gly Arg Arg Phe Leu Leu Leu Gly Thr 595 600 605 Ile Pro Ile Leu Ile Ala Ser Leu Val Ile Leu Val Val Ser Asn Leu 610 615 620 Ile Asp Leu Gly Thr Leu Ala His Ala Leu Leu Ser Thr Ile Ser Val 625 630 635 640 Ile Val Tyr Phe Cys Cys Phe Val Met Gly Phe Gly Pro Ile Pro Asn 645 650 655 Ile Leu Cys Ala Glu Ile Phe Pro Thr Arg Val Arg Gly Leu Cys Ile 660 665 670 Ala Ile Cys Ala Phe Thr Phe Trp Ile Gly Asp Ile Ile Val Thr Tyr 675 680 685 Ser Leu Pro Val Met Leu Asn Ala Ile Gly Leu Ala Gly Val Phe Ser 690 695 700 Ile Tyr Ala Val Val Cys Leu Ile Ser Phe Val Phe Val Phe Leu Lys 705 710 715 720 Val Pro Glu Thr Lys Gly Met Pro Leu Glu Val Ile Thr Glu Phe Phe 725 730 735 Ala Val Gly Ala Lys Gln Ala Ala Ala Lys Ala 740 745 3 443 DNA Oryza sativa unsure (193) unsure (388) unsure (435) unsure (439) 3 gaagagctca cccccccccc ctcggccctg gactccctcc tccaaatctc ccctaaaagc 60 ttcccaattt ggcgagaatt ccccatatat ttgccccatc tcggcgtccc aacgagccct 120 tccagattcc cagccgcctc tcttcttgtt aggggatccg aaatctcggt ggacgagaga 180 cttggtggta atnattcgcc ggccatggcg ggcgccgtgc tggtcgccat cgcggcctcc 240 atcggcaact tgctgcaggg ctgggataat gcaaccattg caggtgcggt actgtacatc 300 aagaaggaat tcaacttgca tagcgacccc cttatcgaag gtctgatcgt ggccatgtcg 360 ctcattgggg ccaccatcat cacgacgntc tctgcgagca ggtggctgac tcttttggta 420 tggcggccca tgctnatcnc ttc 443 4 131 PRT Oryza sativa UNSURE (65) UNSURE (130) 4 Glu Glu Leu Thr Pro Pro Pro Ser Ala Leu Asp Ser Leu Leu Gln Ile 1 5 10 15 Ser Pro Lys Ser Phe Pro Ile Trp Arg Glu Phe Pro Ile Tyr Leu Pro 20 25 30 His Leu Gly Val Pro Thr Ser Pro Ser Arg Phe Pro Ala Ala Ser Leu 35 40 45 Leu Val Arg Gly Ser Glu Ile Ser Val Asp Glu Arg Leu Gly Gly Asn 50 55 60 Xaa Ser Pro Ala Met Ala Gly Ala Val Leu Val Ala Ile Ala Ala Ser 65 70 75 80 Ile Gly Asn Leu Leu Gln Gly Trp Asp Asn Ala Thr Ile Ala Gly Ala 85 90 95 Val Leu Tyr Ile Lys Lys Glu Phe Asn Leu His Ser Asp Pro Leu Ile 100 105 110 Glu Gly Leu Ile Val Ala Met Ser Leu Ile Gly Ala Thr Ile Ile Thr 115 120 125 Thr Xaa Ser 130 5 870 DNA Oryza sativa 5 gcacgaggtt ctaaccttga ttctggtcaa tattctggat gtggggacca tggttcatgc 60 ctcactgtcc acagtcagtg tcatactcta cttctgcttc tttgtcatgg ggttcgggcc 120 tattccaaac attctctgtg cagagatttt cccgaccacc gttcgtggca tctgcatagc 180 catctgtgcc ctaacattct ggatcggtga tatcattgtg acatacaccc tccccgtgat 240 gctcaacgcc attggactcg ctggagtgtt tggaatctac gcagtggtct gcatactggc 300 tttcctgttt gtcttcatga aggtgccgga gacaaagggc atgcctcttg aagtcatcac 360 cgagttcttc tctgtcggag caaagcaggc caaggaggac tagttgctcg gatcaagtga 420 tcaatcagat tgctggtggt aattttgttg cttccaaatc gcgctgcggg ttaaacctgt 480 gatggatgct ttgttaaaga atcttggaag agatcaaaat gcagtgagcc taaagagatg 540 atttggctgt acatcatgag gctgaatcct gtcgtagact ggattttgga gcttaggata 600 tgtagatcat ctgttccttt tggtttggtc attttccatt tgtgtttctt tggaattctt 660 ctccctgtaa ctagtggtct atcacagttg tgttactggt tttgccttac tcttgagttt 720 gttttcttct ctcggttgtg agttctgaat attagcatag ccgagtacta gttctgaatt 780 ggtttcctct ctgctgaaca tctttcattg atgcttggat ttcatcaaaa aaaaaaaaaa 840 aaaactcgag ggggagcccg gtacacatct 870 6 131 PRT Oryza sativa 6 Val Leu Thr Leu Ile Leu Val Asn Ile Leu Asp Val Gly Thr Met Val 1 5 10 15 His Ala Ser Leu Ser Thr Val Ser Val Ile Leu Tyr Phe Cys Phe Phe 20 25 30 Val Met Gly Phe Gly Pro Ile Pro Asn Ile Leu Cys Ala Glu Ile Phe 35 40 45 Pro Thr Thr Val Arg Gly Ile Cys Ile Ala Ile Cys Ala Leu Thr Phe 50 55 60 Trp Ile Gly Asp Ile Ile Val Thr Tyr Thr Leu Pro Val Met Leu Asn 65 70 75 80 Ala Ile Gly Leu Ala Gly Val Phe Gly Ile Tyr Ala Val Val Cys Ile 85 90 95 Leu Ala Phe Leu Phe Val Phe Met Lys Val Pro Glu Thr Lys Gly Met 100 105 110 Pro Leu Glu Val Ile Thr Glu Phe Phe Ser Val Gly Ala Lys Gln Ala 115 120 125 Lys Glu Asp 130 7 2601 DNA Glycine max 7 gttgcttaac ccttgttgag tgaagtgagc aaggggaatg gcgatctgaa attcggatac 60 tttaattgct tctcgctttc accgaccgaa ctcaatttat agatactccg tcaacctcaa 120 tcccaactaa ctagcagttc cttgctgctg ctccttcttc accatatcgc agtaatgaaa 180 ggtgccgtcc ttgttgctat tgccgcttcc attggtaatt tcctccaagg atgggataat 240 gctaccatcg ccggggctaa tggttacatt aagaaagacc ttgctttggg aacaactatg 300 gaaaggcttg tggtgggcat gtccctgatt ggagcaacgg taatcaccac atgctctggt 360 cctatagcgg attggctcgg tcggcgaccc atgatgataa tctcatctgt gctctatttc 420 ttgggtggtt tggtgatgct gtggtcccca aatgtgtatg tgttgtgctt ggcgaggcta 480 cttgatggat ttgggattgg ccttgctgtg actcttgtcc cggtctatat atctgaaacg 540 gcgccgtctg aaataagggg gtcgttgaat acgcttcctc agttcagtgg ctctggagga 600 atgtttttgt cgtactgtat ggtttttggc atgtcattga gtcccgcgcc tagctggagg 660 ctcatgcttg gggttctgtc tattccttct ctcttgtatt ttgcattgac catttttttc 720 ttgcccgagt ctcctcggtg gctggtcagc aaaggaagga tgctcgaggc taagaaggtg 780 ctccaaagat tgcgcggaag ggaggatgtg tcaggcgaga tggcattgct ggttgaaggt 840 ctcgggattg ggggtgatac atctatcgaa gagtacataa ttggccctgc tgacgatgtg 900 gctgatggtc atgaacatgc aacagagaaa gataaaattc gattatatgg atcccaagca 960 ggcctttctt ggttatcaaa acctgtcact ggacagagtt ctattggcct tgcgtcacac 1020 catggaagca tcatcaacca aagcatgccc ctcatggatc ctctggtgac actgtttggt 1080 agcattcatg agaagctccc cgagacagga gcaagaggaa gcatgcgaag cactctgttt 1140 ccaaattttg gaagcatgtt cagcactgct gagccgcatg ctaaaattga acaatgggat 1200 gaagaaagct tacaaaggga acgtgaggac tacatgtcag atgcaacccg tggggactcc 1260 gatgataatt tgcacagtcc tttaatctca cgccaaacaa caagccttga aaaagactta 1320 cctcctcctc cttcccatgg cagtatcctt ggcagcatga ggcgtcacag tagtctcatg 1380 caagggtcag gtgagcaagg tggtagtaca ggtattggtg gtggctggca actggcatgg 1440 aaatggactg ataaaggtga ggatggaaaa caacaaggag ggtttaaaag gatttattta 1500 catgaggagg gagtttctgc atctcgtcgt ggatccattg tatcgattcc cggtgaaggc 1560 gaatttgtcc aggctgctgc cttggtaagc caacccgctc tttactccaa ggagcttatt 1620 gatggacacc cagttgggcc tgcaatggtt cacccatctg agacagcttc aaaggggcca 1680 agttggaaag ctcttcttga accaggggtt aagcatgcat tggttgttgg agttggaata 1740 caaatacttc agcagttttc agggataaat ggggttctat attacacacc tcaaatcctt 1800 gaagaggccg gtgttgaagt tcttctttca gatataggca ttggctcaga gtcggcatca 1860 ttccttatca gtgctttcac aaccttcttg atgcttccct gtataggcgt agccatgaag 1920 ctcatggatg tttcaggcag aaggcagttg ctacttacta caatccccgt gctgattgtg 1980 tcactcatta ttttggtcat tggaagcctg gtaaattttg gcaatgtcgc ccatgcagca 2040 atctcaacag tatgcgttgt ggtttatttc tgctgctttg tgatgggtta tggaccaatt 2100 ccaaacatcc tttgctcaga gattttcccc actagggtgc gtggcctctg cattgctatc 2160 tgtgcattag tgttctggat tggagacatc atcatcacat actcgctgcc tgtgatgctc 2220 ggctctttag gacttggtgg tgtattcgcc atttacgcag ttgtttgttt catctcgtgg 2280 atatttgtgt ttttgaaggt tccagaaaca aagggcatgc cccttgaagt catctctgaa 2340 ttcttttctg ttggagcaaa gcaggctgct tctgccaaga atgagtgaca caacacaagt 2400 ccgttatata ctctgtaact ttagttgtta aagccatcat ctctcgtctt tacagatttt 2460 gcttttcata agtttatttg gaggaagata ttttgaaaca tatgggtttt tttttctttc 2520 ataaaaataa aacccttccc tttttgggtg gggaaaagaa aaaaaaaaaa aaaaaaaaaa 2580 aaaaaaaaaa aaaaaaaaaa a 2601 8 737 PRT Glycine max 8 Met Lys Gly Ala Val Leu Val Ala Ile Ala Ala Ser Ile Gly Asn Phe 1 5 10 15 Leu Gln Gly Trp Asp Asn Ala Thr Ile Ala Gly Ala Asn Gly Tyr Ile 20 25 30 Lys Lys Asp Leu Ala Leu Gly Thr Thr Met Glu Arg Leu Val Val Gly 35 40 45 Met Ser Leu Ile Gly Ala Thr Val Ile Thr Thr Cys Ser Gly Pro Ile 50 55 60 Ala Asp Trp Leu Gly Arg Arg Pro Met Met Ile Ile Ser Ser Val Leu 65 70 75 80 Tyr Phe Leu Gly Gly Leu Val Met Leu Trp Ser Pro Asn Val Tyr Val 85 90 95 Leu Cys Leu Ala Arg Leu Leu Asp Gly Phe Gly Ile Gly Leu Ala Val 100 105 110 Thr Leu Val Pro Val Tyr Ile Ser Glu Thr Ala Pro Ser Glu Ile Arg 115 120 125 Gly Ser Leu Asn Thr Leu Pro Gln Phe Ser Gly Ser Gly Gly Met Phe 130 135 140 Leu Ser Tyr Cys Met Val Phe Gly Met Ser Leu Ser Pro Ala Pro Ser 145 150 155 160 Trp Arg Leu Met Leu Gly Val Leu Ser Ile Pro Ser Leu Leu Tyr Phe 165 170 175 Ala Leu Thr Ile Phe Phe Leu Pro Glu Ser Pro Arg Trp Leu Val Ser 180 185 190 Lys Gly Arg Met Leu Glu Ala Lys Lys Val Leu Gln Arg Leu Arg Gly 195 200 205 Arg Glu Asp Val Ser Gly Glu Met Ala Leu Leu Val Glu Gly Leu Gly 210 215 220 Ile Gly Gly Asp Thr Ser Ile Glu Glu Tyr Ile Ile Gly Pro Ala Asp 225 230 235 240 Asp Val Ala Asp Gly His Glu His Ala Thr Glu Lys Asp Lys Ile Arg 245 250 255 Leu Tyr Gly Ser Gln Ala Gly Leu Ser Trp Leu Ser Lys Pro Val Thr 260 265 270 Gly Gln Ser Ser Ile Gly Leu Ala Ser His His Gly Ser Ile Ile Asn 275 280 285 Gln Ser Met Pro Leu Met Asp Pro Leu Val Thr Leu Phe Gly Ser Ile 290 295 300 His Glu Lys Leu Pro Glu Thr Gly Ala Arg Gly Ser Met Arg Ser Thr 305 310 315 320 Leu Phe Pro Asn Phe Gly Ser Met Phe Ser Thr Ala Glu Pro His Ala 325 330 335 Lys Ile Glu Gln Trp Asp Glu Glu Ser Leu Gln Arg Glu Arg Glu Asp 340 345 350 Tyr Met Ser Asp Ala Thr Arg Gly Asp Ser Asp Asp Asn Leu His Ser 355 360 365 Pro Leu Ile Ser Arg Gln Thr Thr Ser Leu Glu Lys Asp Leu Pro Pro 370 375 380 Pro Pro Ser His Gly Ser Ile Leu Gly Ser Met Arg Arg His Ser Ser 385 390 395 400 Leu Met Gln Gly Ser Gly Glu Gln Gly Gly Ser Thr Gly Ile Gly Gly 405 410 415 Gly Trp Gln Leu Ala Trp Lys Trp Thr Asp Lys Gly Glu Asp Gly Lys 420 425 430 Gln Gln Gly Gly Phe Lys Arg Ile Tyr Leu His Glu Glu Gly Val Ser 435 440 445 Ala Ser Arg Arg Gly Ser Ile Val Ser Ile Pro Gly Glu Gly Glu Phe 450 455 460 Val Gln Ala Ala Ala Leu Val Ser Gln Pro Ala Leu Tyr Ser Lys Glu 465 470 475 480 Leu Ile Asp Gly His Pro Val Gly Pro Ala Met Val His Pro Ser Glu 485 490 495 Thr Ala Ser Lys Gly Pro Ser Trp Lys Ala Leu Leu Glu Pro Gly Val 500 505 510 Lys His Ala Leu Val Val Gly Val Gly Ile Gln Ile Leu Gln Gln Phe 515 520 525 Ser Gly Ile Asn Gly Val Leu Tyr Tyr Thr Pro Gln Ile Leu Glu Glu 530 535 540 Ala Gly Val Glu Val Leu Leu Ser Asp Ile Gly Ile Gly Ser Glu Ser 545 550 555 560 Ala Ser Phe Leu Ile Ser Ala Phe Thr Thr Phe Leu Met Leu Pro Cys 565 570 575 Ile Gly Val Ala Met Lys Leu Met Asp Val Ser Gly Arg Arg Gln Leu 580 585 590 Leu Leu Thr Thr Ile Pro Val Leu Ile Val Ser Leu Ile Ile Leu Val 595 600 605 Ile Gly Ser Leu Val Asn Phe Gly Asn Val Ala His Ala Ala Ile Ser 610 615 620 Thr Val Cys Val Val Val Tyr Phe Cys Cys Phe Val Met Gly Tyr Gly 625 630 635 640 Pro Ile Pro Asn Ile Leu Cys Ser Glu Ile Phe Pro Thr Arg Val Arg 645 650 655 Gly Leu Cys Ile Ala Ile Cys Ala Leu Val Phe Trp Ile Gly Asp Ile 660 665 670 Ile Ile Thr Tyr Ser Leu Pro Val Met Leu Gly Ser Leu Gly Leu Gly 675 680 685 Gly Val Phe Ala Ile Tyr Ala Val Val Cys Phe Ile Ser Trp Ile Phe 690 695 700 Val Phe Leu Lys Val Pro Glu Thr Lys Gly Met Pro Leu Glu Val Ile 705 710 715 720 Ser Glu Phe Phe Ser Val Gly Ala Lys Gln Ala Ala Ser Ala Lys Asn 725 730 735 Glu 9 1692 DNA Glycine max 9 gcacgaggga tccgtccaga gaaaaagatc aaattaagtt gtatggacca gaacaaggcc 60 agtcctgggt tgctagacct gttgctggac caaattctgt tggccttgta tctaggaaag 120 gaagcatggc aaatccaagc agtctagtgg accctctagt gaccctcttt ggtagtgtac 180 atgagaagct cccagaaaca ggaagcaccc tttttccaca ctttgggagt atgttcagtg 240 ttgggggaaa tcagccaagg aatgaagatt gggatgagga aagcctagcc agagagggtg 300 atgattatgt ctctgatgct ggtgattctg atgacaattt gcagagtcca ttgatctcac 360 gtcaaacaac gagtctggat aaggacatac ctcctcatgc ccatagtaac cttgcaagca 420 tgaggcaagg tagtctttta catggaaatt caggagaacc cactggtagt actgggattg 480 gtggtggttg gcagctagca tggaaatggt ctgaaagaga gggcccagat ggaaagaagg 540 aaggtggctt caagagaata tatttacacc aagatggtgg ttctggatct agacgtgggt 600 ctgtggtttc actccctggc ggtgatttac caactgacag tgaggttgta caggctgctg 660 ctctggtgag tcagcctgcc ctttataatg aggaccttat gcgtcaacgg ccagttggac 720 cagctatgat tcatccctct gaaacaattg caaaagggcc aagttggagt gatctttttg 780 aacctggggt gaagcatgca ttgattgtgg gggtgggaat gcaaattctt cagcagttct 840 ctggtataaa tggggtcctc tactatacgc ctcaaattct tgagcaggca ggtgttggtt 900 atcttctttc aagcctaggc cttggttcta cttcttcatc ctttcttatt agtgcggtga 960 caaccttgtt gatgcttcct tgtatagcca ttgccatgag gctcatggat atttcaggca 1020 gaaggacttt gctgctcagt acaatccccg tcctaatagc agctcttctc atattagtcc 1080 tgggaagtct tgtggatttg ggatccactg caaatgcatc aatctcaacc attagtgtta 1140 ttgtctattt ctgtttcttt gtcatgggat ttggaccaat tcctaatata ctttgtgcag 1200 agatcttccc cactcgagtt cgtggtctct gcattgctat ttgtgccctt accttttgga 1260 tctgtgatat cattgtcacc tacacactcc cagttatgct caattctgta ggcctcgctg 1320 gtgtttttgg tatttatgct gtcgtgtgct tcatagcatg ggtgtttgtc tttttgaaag 1380 ttccagaaac caagggcatg ccactggaag tgatcattga gttcttctct gtcggagcaa 1440 aacagtttga cgatgccaag cacaactgac ccaaggacat gataaattca aagttttgac 1500 ggtaccttct aattattttc aatctacggc tgtttgaaat tttcccctct tttaaaattt 1560 tattttctat ttattctctc ttttccgtgg gttgagattg agaaacaaga aactttgttt 1620 ctgtaaagaa aaatgttcat tttctggttc atttatggaa ctttatatac ttcctaaaaa 1680 aaaaaaaaaa aa 1692 10 486 PRT Glycine max 10 Asp Pro Ser Arg Glu Lys Asp Gln Ile Lys Leu Tyr Gly Pro Glu Gln 1 5 10 15 Gly Gln Ser Trp Val Ala Arg Pro Val Ala Gly Pro Asn Ser Val Gly 20 25 30 Leu Val Ser Arg Lys Gly Ser Met Ala Asn Pro Ser Ser Leu Val Asp 35 40 45 Pro Leu Val Thr Leu Phe Gly Ser Val His Glu Lys Leu Pro Glu Thr 50 55 60 Gly Ser Thr Leu Phe Pro His Phe Gly Ser Met Phe Ser Val Gly Gly 65 70 75 80 Asn Gln Pro Arg Asn Glu Asp Trp Asp Glu Glu Ser Leu Ala Arg Glu 85 90 95 Gly Asp Asp Tyr Val Ser Asp Ala Gly Asp Ser Asp Asp Asn Leu Gln 100 105 110 Ser Pro Leu Ile Ser Arg Gln Thr Thr Ser Leu Asp Lys Asp Ile Pro 115 120 125 Pro His Ala His Ser Asn Leu Ala Ser Met Arg Gln Gly Ser Leu Leu 130 135 140 His Gly Asn Ser Gly Glu Pro Thr Gly Ser Thr Gly Ile Gly Gly Gly 145 150 155 160 Trp Gln Leu Ala Trp Lys Trp Ser Glu Arg Glu Gly Pro Asp Gly Lys 165 170 175 Lys Glu Gly Gly Phe Lys Arg Ile Tyr Leu His Gln Asp Gly Gly Ser 180 185 190 Gly Ser Arg Arg Gly Ser Val Val Ser Leu Pro Gly Gly Asp Leu Pro 195 200 205 Thr Asp Ser Glu Val Val Gln Ala Ala Ala Leu Val Ser Gln Pro Ala 210 215 220 Leu Tyr Asn Glu Asp Leu Met Arg Gln Arg Pro Val Gly Pro Ala Met 225 230 235 240 Ile His Pro Ser Glu Thr Ile Ala Lys Gly Pro Ser Trp Ser Asp Leu 245 250 255 Phe Glu Pro Gly Val Lys His Ala Leu Ile Val Gly Val Gly Met Gln 260 265 270 Ile Leu Gln Gln Phe Ser Gly Ile Asn Gly Val Leu Tyr Tyr Thr Pro 275 280 285 Gln Ile Leu Glu Gln Ala Gly Val Gly Tyr Leu Leu Ser Ser Leu Gly 290 295 300 Leu Gly Ser Thr Ser Ser Ser Phe Leu Ile Ser Ala Val Thr Thr Leu 305 310 315 320 Leu Met Leu Pro Cys Ile Ala Ile Ala Met Arg Leu Met Asp Ile Ser 325 330 335 Gly Arg Arg Thr Leu Leu Leu Ser Thr Ile Pro Val Leu Ile Ala Ala 340 345 350 Leu Leu Ile Leu Val Leu Gly Ser Leu Val Asp Leu Gly Ser Thr Ala 355 360 365 Asn Ala Ser Ile Ser Thr Ile Ser Val Ile Val Tyr Phe Cys Phe Phe 370 375 380 Val Met Gly Phe Gly Pro Ile Pro Asn Ile Leu Cys Ala Glu Ile Phe 385 390 395 400 Pro Thr Arg Val Arg Gly Leu Cys Ile Ala Ile Cys Ala Leu Thr Phe 405 410 415 Trp Ile Cys Asp Ile Ile Val Thr Tyr Thr Leu Pro Val Met Leu Asn 420 425 430 Ser Val Gly Leu Ala Gly Val Phe Gly Ile Tyr Ala Val Val Cys Phe 435 440 445 Ile Ala Trp Val Phe Val Phe Leu Lys Val Pro Glu Thr Lys Gly Met 450 455 460 Pro Leu Glu Val Ile Ile Glu Phe Phe Ser Val Gly Ala Lys Gln Phe 465 470 475 480 Asp Asp Ala Lys His Asn 485 11 510 DNA Triticum aestivum unsure (421) unsure (434) unsure (441) unsure (458) unsure (483) unsure (493) unsure (498) 11 cggtggcagc cggggcagtg aaggaggggt agctcttggc tcctatttga ggcggcttcg 60 ctcggttctg atctaccgca ccacaccacc acaccacacc aggggcctgc cgcttcttgg 120 gcttctccat ctcatctcct tggttggttc tctactagag aggcgcagct gcagggatcc 180 ttggtggaga ggagggaaga agatgtcggg tgctgcactg gtcgcgattg cggcttccat 240 tggcaatctg ctgcaggggt gggacaatgc caccatcgct ggtgctgttc tgtacatcaa 300 gaaggaattc cagctcgaaa ataatccgac tgtggagggg ctcatcgtgg catgtcctca 360 tcgggtgcaa catcatcaca cattctccgg gccagtatca aactgggttg ccgggcccta 420 ngccatctcc ttgntttcaa ntcccaaggg ctaatcanct aggcaccaat gtcaatgtgc 480 gcnccggaac ctntcaangg ttggaacgtt 510 12 117 PRT Triticum aestivum 12 Gly Gly Ser Arg Gly Ser Glu Gly Gly Val Ala Leu Gly Ser Tyr Leu 1 5 10 15 Arg Arg Leu Arg Ser Val Leu Ile Tyr Arg Thr Thr Pro Pro His His 20 25 30 Thr Arg Gly Leu Pro Leu Leu Gly Leu Leu His Leu Ile Ser Leu Val 35 40 45 Gly Ser Leu Leu Glu Arg Arg Ser Cys Arg Asp Pro Trp Trp Arg Gly 50 55 60 Gly Lys Lys Met Ser Gly Ala Ala Leu Val Ala Ile Ala Ala Ser Ile 65 70 75 80 Gly Asn Leu Leu Gln Gly Trp Asp Asn Ala Thr Ile Ala Gly Ala Val 85 90 95 Leu Tyr Ile Lys Lys Glu Phe Gln Leu Glu Asn Asn Pro Thr Val Glu 100 105 110 Gly Leu Ile Val Ala 115 13 1487 DNA Triticum aestivum 13 tctcttggaa agagggtggg gaggcagtca gcagcactgg tattggtggg gggtggcaac 60 tcgcatggaa atggtcggag cgacaaggcg aggatggcaa gaaggaagga ggcttcaaaa 120 gaatctactt gcaccaagag ggggtggccg actcaagaag gggctctgtt gtttcacttc 180 ctggtggggg tgatgccacg caagggggca gtgggtttat acatgctgct gctttggtaa 240 gccactcggc tctttactcc aaggatctta tggaagagcg tatggcggcc ggtccagcca 300 tgattcatcc attggaggca gctcccaaag gttcaatctg gaaagatctg tttgaacctg 360 gtgtgaggcg tgcattgttc gtcggtgttg gaattcagat gcttcagcag tttgctggaa 420 taaatggagt tctctactat actcctcaaa ttctggagca agctggtgtg gctgttcttc 480 tttccaatct tggcctcagt tcagcatcag catccatctt gatcagttct ctcaccacct 540 tactcatgct cccaagcatt ggtgtagcca tgagacttat ggatatatct ggaagaaggt 600 ttctgctact gggcacaatt cccatcttga tagcatccct aattgttttg ggtgtggtca 660 atgttatcaa cttgagtacg gtgccccacg ctgtgctctc cacagttagc gtcattgtct 720 acttctgctg ctttgtcatg ggctttggcc cgatccccaa cattctatgt gcagagattt 780 tccccaccag agtccgtggt gtctgcatcg ctatttgcgc cctcacattc tggatttgtg 840 acattattgt tacctacagc ctgcctgtga tgctgaatgc tattggtcta gcgggtgtct 900 ttggtatata tgcagtcgtt tgctgcattg cctttgtgtt cgtctaccta aaggtcccag 960 agacaaaggg catgcccctc gaggtcatca ccgagttctt tgcggttggg gcgaagcaag 1020 cgcaggccac cattgcctga ttcatcatgg agctttgttt tcagtttgca cactgcggtc 1080 tgcgctgaaa attgcaaatt ggacgggtcc tcgtgaggaa cggaaaaact tttgagttgt 1140 aaatgagaca gctacccaaa gagctcatca cgaggaacgg gaagctgtaa aagtaggagg 1200 atctcatgcc cccatttcat cgtctattat tgcttattag tactgtactg taatcgtcat 1260 tagttgctgt agggttgttc aacttgctaa tctgattctg aactaccatg ctgatgtccg 1320 aaataaagaa aaagcatgtt tttttttgtg tcaacttgca aactttcttt taaacattgt 1380 gcaatgtatt gtaaatttct ttatcaactt ccctcgattc agagagaagc acttgtttgt 1440 aagtcatgaa agatttttct cgacaaaaaa aaaaaaaaaa aaaaaaa 1487 14 345 PRT Triticum aestivum 14 Ser Trp Lys Glu Gly Gly Glu Ala Val Ser Ser Thr Gly Ile Gly Gly 1 5 10 15 Gly Trp Gln Leu Ala Trp Lys Trp Ser Glu Arg Gln Gly Glu Asp Gly 20 25 30 Lys Lys Glu Gly Gly Phe Lys Arg Ile Tyr Leu His Gln Glu Gly Val 35 40 45 Ala Asp Ser Arg Arg Gly Ser Val Val Ser Leu Pro Gly Gly Gly Asp 50 55 60 Ala Thr Gln Gly Gly Ser Gly Phe Ile His Ala Ala Ala Leu Val Ser 65 70 75 80 His Ser Ala Leu Tyr Ser Lys Asp Leu Met Glu Glu Arg Met Ala Ala 85 90 95 Gly Pro Ala Met Ile His Pro Leu Glu Ala Ala Pro Lys Gly Ser Ile 100 105 110 Trp Lys Asp Leu Phe Glu Pro Gly Val Arg Arg Ala Leu Phe Val Gly 115 120 125 Val Gly Ile Gln Met Leu Gln Gln Phe Ala Gly Ile Asn Gly Val Leu 130 135 140 Tyr Tyr Thr Pro Gln Ile Leu Glu Gln Ala Gly Val Ala Val Leu Leu 145 150 155 160 Ser Asn Leu Gly Leu Ser Ser Ala Ser Ala Ser Ile Leu Ile Ser Ser 165 170 175 Leu Thr Thr Leu Leu Met Leu Pro Ser Ile Gly Val Ala Met Arg Leu 180 185 190 Met Asp Ile Ser Gly Arg Arg Phe Leu Leu Leu Gly Thr Ile Pro Ile 195 200 205 Leu Ile Ala Ser Leu Ile Val Leu Gly Val Val Asn Val Ile Asn Leu 210 215 220 Ser Thr Val Pro His Ala Val Leu Ser Thr Val Ser Val Ile Val Tyr 225 230 235 240 Phe Cys Cys Phe Val Met Gly Phe Gly Pro Ile Pro Asn Ile Leu Cys 245 250 255 Ala Glu Ile Phe Pro Thr Arg Val Arg Gly Val Cys Ile Ala Ile Cys 260 265 270 Ala Leu Thr Phe Trp Ile Cys Asp Ile Ile Val Thr Tyr Ser Leu Pro 275 280 285 Val Met Leu Asn Ala Ile Gly Leu Ala Gly Val Phe Gly Ile Tyr Ala 290 295 300 Val Val Cys Cys Ile Ala Phe Val Phe Val Tyr Leu Lys Val Pro Glu 305 310 315 320 Thr Lys Gly Met Pro Leu Glu Val Ile Thr Glu Phe Phe Ala Val Gly 325 330 335 Ala Lys Gln Ala Gln Ala Thr Ile Ala 340 345 15 1009 DNA Triticum aestivum 15 tgaacctgga gtgaagcatg cactgttcgt tggcatagga ttacagatcc tgcagcagtt 60 tgcgggtatc aatggagtcc tctactacac acctcagata cttgagcaag caggtgtcgg 120 ggttcttcta tcaaacattg gactaagctc ttcctcagca tctattctta ttagtgcctt 180 gacaaccttg ctgatgcttc ccagcattgg catcgccatg agactcatgg atatgtcagg 240 aagaaggttt cttctccttt caacaatccc tgtcttgata gtagcgctag ctgtcttggt 300 tttagtgaat gttctggatg tcggaaccat ggtgcacgct gcgctctcaa cgatcagcgt 360 catcgtctat ttctgcttct tcgtcatggg gtttgggcct atcccaaata ttctctgcgc 420 ggagattttc cccacctctg tccgtggcat ctgcatagcc atctgcgcgc taaccttctg 480 gatcggcgac atcatcgtga catacactct ccccgtgatg ctcaatgcca ttggtctcgc 540 tggagtcttc ggcatatatg ccatcgtttg tgtactagcc tttgtattcg tctacatgaa 600 ggtccctgag acaaagggca tgcccctgga ggtcatcacc gagttcttct ctgtcggggc 660 aaagcagggc aaggaagcca cggactagtt gctctgatcc ggtgatccgc gtcgctggtg 720 gtaattttgt ggtgtcataa ctactactac actggttaac ctgcgatgct ttggtgaaga 780 aacttcaaag agagcagata cggaagactt tacatcgtga ggctgaattg tgtcgtcgta 840 ggccggcttt tggaagtagg atatgtactt agatcatctg ctcttttcgc tttggaactt 900 tctatttgtg ttattcagaa tttcttgccc atgtaactag tgctgttatc acaatttatg 960 tcgattatgt gtttgcctaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaa 1009 16 228 PRT Triticum aestivum 16 Glu Pro Gly Val Lys His Ala Leu Phe Val Gly Ile Gly Leu Gln Ile 1 5 10 15 Leu Gln Gln Phe Ala Gly Ile Asn Gly Val Leu Tyr Tyr Thr Pro Gln 20 25 30 Ile Leu Glu Gln Ala Gly Val Gly Val Leu Leu Ser Asn Ile Gly Leu 35 40 45 Ser Ser Ser Ser Ala Ser Ile Leu Ile Ser Ala Leu Thr Thr Leu Leu 50 55 60 Met Leu Pro Ser Ile Gly Ile Ala Met Arg Leu Met Asp Met Ser Gly 65 70 75 80 Arg Arg Phe Leu Leu Leu Ser Thr Ile Pro Val Leu Ile Val Ala Leu 85 90 95 Ala Val Leu Val Leu Val Asn Val Leu Asp Val Gly Thr Met Val His 100 105 110 Ala Ala Leu Ser Thr Ile Ser Val Ile Val Tyr Phe Cys Phe Phe Val 115 120 125 Met Gly Phe Gly Pro Ile Pro Asn Ile Leu Cys Ala Glu Ile Phe Pro 130 135 140 Thr Ser Val Arg Gly Ile Cys Ile Ala Ile Cys Ala Leu Thr Phe Trp 145 150 155 160 Ile Gly Asp Ile Ile Val Thr Tyr Thr Leu Pro Val Met Leu Asn Ala 165 170 175 Ile Gly Leu Ala Gly Val Phe Gly Ile Tyr Ala Ile Val Cys Val Leu 180 185 190 Ala Phe Val Phe Val Tyr Met Lys Val Pro Glu Thr Lys Gly Met Pro 195 200 205 Leu Glu Val Ile Thr Glu Phe Phe Ser Val Gly Ala Lys Gln Gly Lys 210 215 220 Glu Ala Thr Asp 225 17 615 DNA Zea mays unsure (149) unsure (271) unsure (304) unsure (334) unsure (357) unsure (476) unsure (599) unsure (602) 17 gaaacgaact ctcttgagta ccacaaaaaa aaacattggc attctctgta gtagagcaca 60 gagcgaaccg tcaacgatgg cttccgctcc gctgccggcg gccatcgagc ccgggaagaa 120 aggcaacgtc aagttcgcct tcgcctgcnc catcctcgcc tcaatgacct ccatccttct 180 cggctatgat atcggagtga tgagcggcgc gtcgttgtac atcaagaagg acctgaaaat 240 cagcgacgtg aagctggaga tcctgatggg natcctcaac gtgtactcgc tcatcggctc 300 gttngcggca gggcggacgt ccgactggat cggncgccgt acaccatcgt gttcgcngcg 360 gtgatcttct tcgcgggcgc ttcctcatgg gcttcgccgt gaactactgg atgctcatgt 420 tcgggcgctt cgtggccggg atcggcgtgg gctacgcgct catgatcgca accgtntaca 480 cggccgaagt gtccccgcat cggcccgcgg cttcctgacg tcgttcccgg aggtgttcat 540 cacttcggca tcctctaggt acgtgtcaat aaggcttttc cgcttccgtt cgctggatng 600 cnctaatgtc ggcat 615 18 167 PRT Zea mays UNSURE (34) UNSURE (85) UNSURE (98) UNSURE (112) UNSURE (151) 18 Ser Arg Ala Gln Ser Glu Pro Ser Thr Met Ala Ser Ala Pro Leu Pro 1 5 10 15 Ala Ala Ile Glu Pro Gly Lys Lys Gly Asn Val Lys Phe Ala Phe Ala 20 25 30 Cys Xaa Ile Leu Ala Ser Met Thr Ser Ile Leu Leu Gly Tyr Asp Ile 35 40 45 Gly Val Met Ser Gly Ala Ser Leu Tyr Ile Lys Lys Asp Leu Lys Ile 50 55 60 Ser Asp Val Lys Leu Glu Ile Leu Met Gly Ile Leu Asn Val Tyr Ser 65 70 75 80 Leu Ile Gly Ser Xaa Ala Ala Gly Arg Thr Ser Asp Trp Ile Gly Arg 85 90 95 Arg Xaa Thr Ile Val Phe Ala Ala Val Ile Phe Phe Ala Gly Ala Xaa 100 105 110 Leu Met Gly Phe Ala Val Asn Tyr Trp Met Leu Met Phe Gly Arg Phe 115 120 125 Val Ala Gly Ile Gly Val Gly Tyr Ala Leu Met Ile Ala Thr Val Tyr 130 135 140 Thr Ala Glu Val Ser Pro Xaa Ser Ala Arg Gly Phe Leu Thr Ser Phe 145 150 155 160 Pro Glu Val Phe Ile Thr Ser 165 19 1914 DNA Zea mays 19 gcacgaggca cgccacctta tctctaaccg gagatcaaag aagtagccgt taacgatggc 60 ttccgacgag ctcgcaaagg ccgtcgagcc caggaagaag ggcaacgtca agtatgcctc 120 catatgtgcc atcctggcct ccatggcctc tgtcatcctt ggctatgaca ttggggtgat 180 gagtggagcg gccatgtaca tcaagaagga cctgaatatc acggacgtgc agctggagat 240 cctgatcggg atcctcagtc tctactcgct gttcggatcc ttcgctggcg cgcggacgtc 300 cgacaggatc gggcgccgct tgaccgtcgt gttcgccgct gtcatcttct tcgtgggctc 360 gttgctcatg ggtttcgccg tcaactacgg catgctcatg gcgggccgct tcgtggccgg 420 agtcggtgtg ggctacgggg gcatgatcgc gcccgtgtac acggccgaga tctcgcctgc 480 ggcgtcccgt ggcttcctga ccaccttccc ggaggtgttc atcaacatcg gcatcctgct 540 tggctacctg tccaacttcg cgttcgcgcg cctcccgctc cacctcggct ggcgcgtcat 600 gctcgccatt ggcgcagttc cgtccggcct gctcgcgctc ctggtgttct gcatgcccga 660 gtcgcctcgg tggctggtct tgaagggccg cctcgcggac gccagggctg tgctagagaa 720 gacctctgcc acgccagagg aggccgccga gcggctggcc gacatcaagg ccgcggcggg 780 gattccgaag ggcctcgacg gggacgtagt caccgtaccc ggcaaggagc aaggcggcgg 840 tgagttgcag gtgtggaaga agctcatcct gtccccgacc ccggctgtcc gacgcatact 900 gctctcggcc gtgggtctcc acttcttcca gcaggcttct ggcagcgact ccgtcgtcca 960 gtacagcgcc cgcctgttca agagcgcggg gatcaccgac gacaacaagc tcctgggcgt 1020 cacctgcgcg gtgggcgtga ccaagacgtt cttcatcctg gtggccacgt tcctgctgga 1080 ccgcgcgggg cgtcggcctc tgctgctgat cagcacgggc gggatgattg tctcgctcat 1140 ctgcctcggg tcggggctca ccgtcgcggg gcatcacccg gacaccaagg tcgcgtgggc 1200 cgtcgccctg tgcatcgcgt caaccctgtc ctacatcgcc ttcttctcca tcggcctcgg 1260 gcccatcacg ggcgtgtaca cctcggaaat attcccgctg caggtgcgcg cgctgggctt 1320 cgcggtgggt gtggcgagca accgcgtcac cagcgccgtc atctccatga ccttcctgtc 1380 cctctccaag gccatcacca tcggcggcag cttcttcctc tactccggca tcgccgcggt 1440 cgcttgggtt ttcttcttca cgtgcctccc ggagacacgc ggccggacgc tggaggagat 1500 gggcaagctg ttcggcatgc cagacacggg catggctgaa gaagcagaag acgccgcagc 1560 caaggagaag gtggtggaac tgcctagcag caagtaggtg gctatcccag agcacaggtc 1620 aagtgaagta gatggacaag atcattgtct tttcaactaa ttagatgggc aagaataact 1680 aagactgccc tatgaggtgt cgtggttcaa ccagagatca ttctgctcct tttcttttcc 1740 cttccttttt cgagtaccat tcccattcgt cgtggtcagt acgatgttgg gtcgttggga 1800 gttagtggtg tcagagtccg cgtgtgcttt gcaagccagg gctgaaccca caatcatcag 1860 taacaaaaat tcttccgttt gctttgcaag ccaaaaaaaa aaaaaaaaaa aaaa 1914 20 513 PRT Zea mays 20 Met Ala Ser Asp Glu Leu Ala Lys Ala Val Glu Pro Arg Lys Lys Gly 1 5 10 15 Asn Val Lys Tyr Ala Ser Ile Cys Ala Ile Leu Ala Ser Met Ala Ser 20 25 30 Val Ile Leu Gly Tyr Asp Ile Gly Val Met Ser Gly Ala Ala Met Tyr 35 40 45 Ile Lys Lys Asp Leu Asn Ile Thr Asp Val Gln Leu Glu Ile Leu Ile 50 55 60 Gly Ile Leu Ser Leu Tyr Ser Leu Phe Gly Ser Phe Ala Gly Ala Arg 65 70 75 80 Thr Ser Asp Arg Ile Gly Arg Arg Leu Thr Val Val Phe Ala Ala Val 85 90 95 Ile Phe Phe Val Gly Ser Leu Leu Met Gly Phe Ala Val Asn Tyr Gly 100 105 110 Met Leu Met Ala Gly Arg Phe Val Ala Gly Val Gly Val Gly Tyr Gly 115 120 125 Gly Met Ile Ala Pro Val Tyr Thr Ala Glu Ile Ser Pro Ala Ala Ser 130 135 140 Arg Gly Phe Leu Thr Thr Phe Pro Glu Val Phe Ile Asn Ile Gly Ile 145 150 155 160 Leu Leu Gly Tyr Leu Ser Asn Phe Ala Phe Ala Arg Leu Pro Leu His 165 170 175 Leu Gly Trp Arg Val Met Leu Ala Ile Gly Ala Val Pro Ser Gly Leu 180 185 190 Leu Ala Leu Leu Val Phe Cys Met Pro Glu Ser Pro Arg Trp Leu Val 195 200 205 Leu Lys Gly Arg Leu Ala Asp Ala Arg Ala Val Leu Glu Lys Thr Ser 210 215 220 Ala Thr Pro Glu Glu Ala Ala Glu Arg Leu Ala Asp Ile Lys Ala Ala 225 230 235 240 Ala Gly Ile Pro Lys Gly Leu Asp Gly Asp Val Val Thr Val Pro Gly 245 250 255 Lys Glu Gln Gly Gly Gly Glu Leu Gln Val Trp Lys Lys Leu Ile Leu 260 265 270 Ser Pro Thr Pro Ala Val Arg Arg Ile Leu Leu Ser Ala Val Gly Leu 275 280 285 His Phe Phe Gln Gln Ala Ser Gly Ser Asp Ser Val Val Gln Tyr Ser 290 295 300 Ala Arg Leu Phe Lys Ser Ala Gly Ile Thr Asp Asp Asn Lys Leu Leu 305 310 315 320 Gly Val Thr Cys Ala Val Gly Val Thr Lys Thr Phe Phe Ile Leu Val 325 330 335 Ala Thr Phe Leu Leu Asp Arg Ala Gly Arg Arg Pro Leu Leu Leu Ile 340 345 350 Ser Thr Gly Gly Met Ile Val Ser Leu Ile Cys Leu Gly Ser Gly Leu 355 360 365 Thr Val Ala Gly His His Pro Asp Thr Lys Val Ala Trp Ala Val Ala 370 375 380 Leu Cys Ile Ala Ser Thr Leu Ser Tyr Ile Ala Phe Phe Ser Ile Gly 385 390 395 400 Leu Gly Pro Ile Thr Gly Val Tyr Thr Ser Glu Ile Phe Pro Leu Gln 405 410 415 Val Arg Ala Leu Gly Phe Ala Val Gly Val Ala Ser Asn Arg Val Thr 420 425 430 Ser Ala Val Ile Ser Met Thr Phe Leu Ser Leu Ser Lys Ala Ile Thr 435 440 445 Ile Gly Gly Ser Phe Phe Leu Tyr Ser Gly Ile Ala Ala Val Ala Trp 450 455 460 Val Phe Phe Phe Thr Cys Leu Pro Glu Thr Arg Gly Arg Thr Leu Glu 465 470 475 480 Glu Met Gly Lys Leu Phe Gly Met Pro Asp Thr Gly Met Ala Glu Glu 485 490 495 Ala Glu Asp Ala Ala Ala Lys Glu Lys Val Val Glu Leu Pro Ser Ser 500 505 510 Lys 21 2017 DNA Oryza sativa 21 cttacatgta agctcgtgcc ggcacgagct tacactcgac cgccactact gtacacggcc 60 cagagcgagc ctcctcctcc tctgcaccac cggagatggc ttccgccgcg ctgccggagg 120 ccgtcgcgcc gaagaagaag ggcaacgtcc ggttcgcctt cgcctgcgcc atcctcgcct 180 ccatgacctc catcctcctc ggctacgata tcggggtgat gagcggggcg tcgctgtaca 240 tcaagaagga cttcaacatc agtgacggga aggtggaggt tctcatgggc atactgaacc 300 tctactcgct catcggctcc ttcgcggcgg ggcggacgtc ggactggatc ggccggcggt 360 acaccatcgt gttcgccgcc gtcatattct tcgcgggggs gttcctcatg gggttcgccg 420 tcaactacgc catgctcatg ttcggccgct tcgtggccgg catcggcgtg ggctacgcgc 480 tcatgatcgc gccggtgtac accgccgagg tgtcgccggc gtcggcgcgt ggcttcctga 540 cgtcgttccc ggaggtgttc atcaacttcg gcatcctgct cgggtacgtc tcgaactatg 600 ctttctcccg cttgccgctg aacctcgggt ggcgcatcat gctcggcatc ggcgcggcgc 660 cgtccgtgct gctcgcgctc atggtgctcg gcatgccgga gtcgccgcgg tggctggtca 720 tgaagggacg cctcgcggac gccaaggtgg tgctggagaa gacctccgac acggcggagg 780 aggccgcgga gcgcctggcc gacatcaagg ccgccgccgg catccctgag gagctcgacg 840 gcgacgtggt gaccgtcccc aagagaggga gcggaaacga gaagcgggtg tggaaggagc 900 tcatcctgtc cccgaccccg gccatgcggc gcatcctgct gtccgggatc ggcatccact 960 tcttccagca tgcgttgggc attcactccg tcgtcttcta cagccctctc gtgttcaaga 1020 gccccggatt aacgaacgac aaacacttct tgggcaccac ttggccgttc ggtgtcacca 1080 agaggctttt catcttgttg gcgactttct tcatcgacgg cgtcgggcgg cggccgctgt 1140 tgctgggcag cacgggcggg ataatcctct ccctcatcgg cctcggcgcc gggctcaccg 1200 tcgtcggcca gcaccccgac gccaagatac cttgggccat cggcctaagc atcgcctcca 1260 ccctcgccta cgtcgccttc ttctccatcg gccttggccc catcacgtgg gtgtacagct 1320 cggagatctt cccgctccag gtgcgcgcgc tgggctgctc gctcggcgtc gccgccaacc 1380 gcgtcaccag cggcgtcatc tccatgacct tcctgtcgct gtccaaggcc atcaccatcg 1440 gcggcagctt cttcctctac tccggcatcg ccgcgctcgc ctgggtgttc ttctacacct 1500 acctcccgga gacccgcggc cggacgctgg aggagatgag caagctgttc ggcgacacgg 1560 ccgccgcctc ggaatcagac gagccagcca aggagaagaa gaaggtggaa atggccgcca 1620 ctaactgatc aaactaaccg caaaatcacc aaatcctaag ggttttcttg caaaaacgtg 1680 tgctgtactg gctagctagc aagtagtagc agcaacgtgg gaagattcgc tgatccggcg 1740 ttgctggaga gcgacggccg gcgacgacaa agctgagctc cagctcgaga cttcttaaaa 1800 tcatcttcaa gtacatggat tttattttgc tctttgcttt gtccgtaaaa gttgtactat 1860 gcgatgaaga ataccagtat gtagcaaggc tgaggttgtg tgtagctact agaagtgtca 1920 gtcacgttgt tcttgtaaga aatgtttaac tgttaattaa gcagtattgt tgcagtaatc 1980 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 2017 22 510 PRT Oryza sativa UNSURE (102) 22 Met Ala Ser Ala Ala Leu Pro Glu Ala Val Ala Pro Lys Lys Lys Gly 1 5 10 15 Asn Val Arg Phe Ala Phe Ala Cys Ala Ile Leu Ala Ser Met Thr Ser 20 25 30 Ile Leu Leu Gly Tyr Asp Ile Gly Val Met Ser Gly Ala Ser Leu Tyr 35 40 45 Ile Lys Lys Asp Phe Asn Ile Ser Asp Gly Lys Val Glu Val Leu Met 50 55 60 Gly Ile Leu Asn Leu Tyr Ser Leu Ile Gly Ser Phe Ala Ala Gly Arg 65 70 75 80 Thr Ser Asp Trp Ile Gly Arg Arg Tyr Thr Ile Val Phe Ala Ala Val 85 90 95 Ile Phe Phe Ala Gly Xaa Phe Leu Met Gly Phe Ala Val Asn Tyr Ala 100 105 110 Met Leu Met Phe Gly Arg Phe Val Ala Gly Ile Gly Val Gly Tyr Ala 115 120 125 Leu Met Ile Ala Pro Val Tyr Thr Ala Glu Val Ser Pro Ala Ser Ala 130 135 140 Arg Gly Phe Leu Thr Ser Phe Pro Glu Val Phe Ile Asn Phe Gly Ile 145 150 155 160 Leu Leu Gly Tyr Val Ser Asn Tyr Ala Phe Ser Arg Leu Pro Leu Asn 165 170 175 Leu Gly Trp Arg Ile Met Leu Gly Ile Gly Ala Ala Pro Ser Val Leu 180 185 190 Leu Ala Leu Met Val Leu Gly Met Pro Glu Ser Pro Arg Trp Leu Val 195 200 205 Met Lys Gly Arg Leu Ala Asp Ala Lys Val Val Leu Glu Lys Thr Ser 210 215 220 Asp Thr Ala Glu Glu Ala Ala Glu Arg Leu Ala Asp Ile Lys Ala Ala 225 230 235 240 Ala Gly Ile Pro Glu Glu Leu Asp Gly Asp Val Val Thr Val Pro Lys 245 250 255 Arg Gly Ser Gly Asn Glu Lys Arg Val Trp Lys Glu Leu Ile Leu Ser 260 265 270 Pro Thr Pro Ala Met Arg Arg Ile Leu Leu Ser Gly Ile Gly Ile His 275 280 285 Phe Phe Gln His Ala Leu Gly Ile His Ser Val Val Phe Tyr Ser Pro 290 295 300 Leu Val Phe Lys Ser Pro Gly Leu Thr Asn Asp Lys His Phe Leu Gly 305 310 315 320 Thr Thr Trp Pro Phe Gly Val Thr Lys Arg Leu Phe Ile Leu Leu Ala 325 330 335 Thr Phe Phe Ile Asp Gly Val Gly Arg Arg Pro Leu Leu Leu Gly Ser 340 345 350 Thr Gly Gly Ile Ile Leu Ser Leu Ile Gly Leu Gly Ala Gly Leu Thr 355 360 365 Val Val Gly Gln His Pro Asp Ala Lys Ile Pro Trp Ala Ile Gly Leu 370 375 380 Ser Ile Ala Ser Thr Leu Ala Tyr Val Ala Phe Phe Ser Ile Gly Leu 385 390 395 400 Gly Pro Ile Thr Trp Val Tyr Ser Ser Glu Ile Phe Pro Leu Gln Val 405 410 415 Arg Ala Leu Gly Cys Ser Leu Gly Val Ala Ala Asn Arg Val Thr Ser 420 425 430 Gly Val Ile Ser Met Thr Phe Leu Ser Leu Ser Lys Ala Ile Thr Ile 435 440 445 Gly Gly Ser Phe Phe Leu Tyr Ser Gly Ile Ala Ala Leu Ala Trp Val 450 455 460 Phe Phe Tyr Thr Tyr Leu Pro Glu Thr Arg Gly Arg Thr Leu Glu Glu 465 470 475 480 Met Ser Lys Leu Phe Gly Asp Thr Ala Ala Ala Ser Glu Ser Asp Glu 485 490 495 Pro Ala Lys Glu Lys Lys Lys Val Glu Met Ala Ala Thr Asn 500 505 510 23 1853 DNA Glycine max 23 gcacgagagt ttctctcttc acatatcatc atacttagat agtcagatac atcacccaat 60 aattaaatta aatacatgct agcactttaa cagtactcct ttctctaata tctctctcat 120 attttccttt ctgcggatat tcagctaatt aaactaagtc actaagatga ctgagggaaa 180 gctagttgaa gctgcagaag ctcataagac acttcaggat ttcgatcctc caaagaagcg 240 caaaaggaac aagtatgctt ttgcttgtgc tatgctggcc tccatgactt ccatcttgct 300 tggttatgat attggagtga tgagtggagc agccatatac ataaaaaggg acctgaaagt 360 ctcggacgag caaatcgaga tcctgctcgg aatcatcaac ctatactctc tgataggctc 420 atgtctcgcc ggcagaacct ccgactggat aggtccccgt tacacgattg ttttcgccgg 480 caccatcttc tttgtcggag cacttctcat gggtttctcc cccaattatt cctttctcat 540 gtttggccgt ttcgtcgctg gcattggcat cggctacgcc ctcatgatag cccccgtcta 600 caccgccgag gtctccccgg cctcctctcg tggcttcctc acttccttcc ctgaggtatt 660 tattaatgga gggatattaa ttggatacat atcaaactat gcattttcga agctgacact 720 aaaggtggga tggcgaatga tgcttggagt tggtgcaata ccttcggtac tcctaacagt 780 aggagtgttg gcgatgccgg agtccccaag gtggcttgtg atgaggggtc gtttgggaga 840 ggcaagaaaa gtgcttaaca aaacctcaga cagcaaggaa gaggcccaac taaggctagc 900 ggaaatcaaa caagccgcag ggatccccga gagttgcaac gacgacgtcg ttcaggtaaa 960 taaacaaagc aacggtgaag gtgtatggaa agagctcttc ctctatccaa cgcccgcaat 1020 tcgtcacatc gtaatcgctg cccttggtat tcacttcttc caacaagcgt cgggcgtaga 1080 cgccgtcgtt ttgtacagcc ccaggatctt cgaaaaggct gggattacaa acgacacgca 1140 taagcttctt gcaaccgtgg ccgttggatt cgttaagacc gtgttcatct tggcggctac 1200 gtttacgttg gaccgcgtgg gtcgtcgtcc gttgttattg tctagtgtcg gcggcatggt 1260 gctctcgctt ctcacgcttg cgatcagcct cactgttatt gatcattcgg agaggaaatt 1320 aatgtgggcc gttggatcga gcatagccat ggtgttggct tacgtggcca cgttctccat 1380 cggtgcgggt cccatcacgt gggtctatag ttctgagatc ttcccgttga ggctgcgggc 1440 gcarggtgcg gccgcgggag ttgcggtgaa taggaccact agcgcggttg tctcaatgac 1500 ttttctgtcc ctcactagag ccatcactat tggtggagct ttcttccttt attgtggcat 1560 tgctactgtt gggtggatat tcttttacac cgtcttgcct gagacccggg gaaaaacgct 1620 cgaagacatg gaagggtctt ttggtacttt taggtccaaa tccaacgcca gcaaggctgt 1680 agaaaatgag aatgggcaag tagcacaagt ccagctagga accaatgtcc aaacttgaaa 1740 aatgagtatt gggacatcca gtaatagtga agtaatttcg tgattttttt tttgtttttt 1800 actttttaga ctagttcttc aaatcaaaac gagaagttaa agtgaaaaaa aaa 1853 24 523 PRT Glycine max 24 Met Thr Glu Gly Lys Leu Val Glu Ala Ala Glu Ala His Lys Thr Leu 1 5 10 15 Gln Asp Phe Asp Pro Pro Lys Lys Arg Lys Arg Asn Lys Tyr Ala Phe 20 25 30 Ala Cys Ala Met Leu Ala Ser Met Thr Ser Ile Leu Leu Gly Tyr Asp 35 40 45 Ile Gly Val Met Ser Gly Ala Ala Ile Tyr Ile Lys Arg Asp Leu Lys 50 55 60 Val Ser Asp Glu Gln Ile Glu Ile Leu Leu Gly Ile Ile Asn Leu Tyr 65 70 75 80 Ser Leu Ile Gly Ser Cys Leu Ala Gly Arg Thr Ser Asp Trp Ile Gly 85 90 95 Pro Arg Tyr Thr Ile Val Phe Ala Gly Thr Ile Phe Phe Val Gly Ala 100 105 110 Leu Leu Met Gly Phe Ser Pro Asn Tyr Ser Phe Leu Met Phe Gly Arg 115 120 125 Phe Val Ala Gly Ile Gly Ile Gly Tyr Ala Leu Met Ile Ala Pro Val 130 135 140 Tyr Thr Ala Glu Val Ser Pro Ala Ser Ser Arg Gly Phe Leu Thr Ser 145 150 155 160 Phe Pro Glu Val Phe Ile Asn Gly Gly Ile Leu Ile Gly Tyr Ile Ser 165 170 175 Asn Tyr Ala Phe Ser Lys Leu Thr Leu Lys Val Gly Trp Arg Met Met 180 185 190 Leu Gly Val Gly Ala Ile Pro Ser Val Leu Leu Thr Val Gly Val Leu 195 200 205 Ala Met Pro Glu Ser Pro Arg Trp Leu Val Met Arg Gly Arg Leu Gly 210 215 220 Glu Ala Arg Lys Val Leu Asn Lys Thr Ser Asp Ser Lys Glu Glu Ala 225 230 235 240 Gln Leu Arg Leu Ala Glu Ile Lys Gln Ala Ala Gly Ile Pro Glu Ser 245 250 255 Cys Asn Asp Asp Val Val Gln Val Asn Lys Gln Ser Asn Gly Glu Gly 260 265 270 Val Trp Lys Glu Leu Phe Leu Tyr Pro Thr Pro Ala Ile Arg His Ile 275 280 285 Val Ile Ala Ala Leu Gly Ile His Phe Phe Gln Gln Ala Ser Gly Val 290 295 300 Asp Ala Val Val Leu Tyr Ser Pro Arg Ile Phe Glu Lys Ala Gly Ile 305 310 315 320 Thr Asn Asp Thr His Lys Leu Leu Ala Thr Val Ala Val Gly Phe Val 325 330 335 Lys Thr Val Phe Ile Leu Ala Ala Thr Phe Thr Leu Asp Arg Val Gly 340 345 350 Arg Arg Pro Leu Leu Leu Ser Ser Val Gly Gly Met Val Leu Ser Leu 355 360 365 Leu Thr Leu Ala Ile Ser Leu Thr Val Ile Asp His Ser Glu Arg Lys 370 375 380 Leu Met Trp Ala Val Gly Ser Ser Ile Ala Met Val Leu Ala Tyr Val 385 390 395 400 Ala Thr Phe Ser Ile Gly Ala Gly Pro Ile Thr Trp Val Tyr Ser Ser 405 410 415 Glu Ile Phe Pro Leu Arg Leu Arg Ala Gln Gly Ala Ala Ala Gly Val 420 425 430 Ala Val Asn Arg Thr Thr Ser Ala Val Val Ser Met Thr Phe Leu Ser 435 440 445 Leu Thr Arg Ala Ile Thr Ile Gly Gly Ala Phe Phe Leu Tyr Cys Gly 450 455 460 Ile Ala Thr Val Gly Trp Ile Phe Phe Tyr Thr Val Leu Pro Glu Thr 465 470 475 480 Arg Gly Lys Thr Leu Glu Asp Met Glu Gly Ser Phe Gly Thr Phe Arg 485 490 495 Ser Lys Ser Asn Ala Ser Lys Ala Val Glu Asn Glu Asn Gly Gln Val 500 505 510 Ala Gln Val Gln Leu Gly Thr Asn Val Gln Thr 515 520 25 2089 DNA Triticum aestivum 25 agcaccacta aactatacac aaggaggacc tcgtcggcat aatcctcagg cagcgagcag 60 aggggcgtcg tcgacgatgg accgcgccgc actcccggcg gccgtcgagc ccaagaagaa 120 gggcaacgtg aggttcgcct tcgcctgcgc catcctcgcc tccatgacct ccatcctcct 180 cggctacgac atcggcgtga tgagcggagc gtcgctgtac atccagaagg atctgaagat 240 caacgacacc cagctggagg tcctcatggg catcctcaac gtgtactcgc tcattggctc 300 cttcgcggcg gggcggacgt ccgactggat cggccggcgc ttcaccatcg tcttcgccgc 360 cgtcatcttc ttcgcgggcg ccctcatcat gggcttctcc gtcaactacg ccatgctcat 420 gttcgggcgc ttcgtggccg gcatcggcgt ggggtacgct ctcatgatcg cgcccgtgaa 480 cacgggcgag gtgtcccccg cgtctgcccg tggggttctc acatccttcc cggaggtgtt 540 catcaacttc ggcatcctcc tcggatatgt ctccaacttc gccttcgccc gcctctccct 600 ccgcctcggc tggcgcatta tgctcggcat aggcgcggtg ccgtccgtcc tgctcgcgtt 660 catggtgctc ggcatgcccg agtctccccg gtggctcgtc atgaagggcc gtctcgcgga 720 cgccaaggtt gtgcttgcca agacgtccga cacgccggaa gaggccgccg agcgcatcgc 780 cgacattaag actgccgccg gcatccctct gggcctcgac ggcgacgtgg tccccgtgcc 840 caaaaacaaa ggaagcagcg aggagaagcg cgttttgaag gacctcatcc tgtcaccgac 900 catagccatg cgccacatcc tcatcgcggg aatcggcatc cacttcttcc agcagtcttc 960 gggcatcgac gccgtcgtgc tctacagccc gctagttttc aagagcgccg gcatcacggg 1020 cgacagccgt ctccgcggca ccaccgtggc ggtcggggcc accaatacgg tcttcatcct 1080 ggtggccacc ttcctcctcg accgcatccg ccggcggccg ctggtgctga ccagcacggg 1140 cggcatgctc gtctccttag tgggcctcgc gacggggctc accgtcatca gccgccaccc 1200 ggacgagaag atcacctggg ccatcgtcct gtgcatcttc tgcatcatgg cctacgtggc 1260 cttcttctcc atcggcctcg gccccatcac gtgggtgtac agctcggaga tcttcccgct 1320 gcacgtgcgc gcgctgggct gctccctggg cgtggccgtc aaccgcctga ccagcggcgt 1380 gatctccatg accttcattt cgctgtccaa ggccatgacc atcggcggcg ccttcttcct 1440 cttcgccggc atcgcctcat tcgcatgggt gttcttcttc gcctacctgc cggagacccg 1500 cggccgcacg ctggaggaca tgagctcgct gttcggcaac acggccacgc acaagcaggg 1560 cgccgcggaa gccgacgacg acgccgggga gaagaaggtg gaaatggccg ccaccaactg 1620 accgcaagtt ggcagatcgc gatgcgaaga cttgcgctgt atccgtctcg gctagctagc 1680 tgccacaagg ccacatagat gacgaagtag cgtgggaaga ttcgctgatc cggccggagc 1740 tgccggaggg cgacggcaag ctccagctcg atcgagacgt taatggcttc ttaaatgtgc 1800 taagtttaat gtttcgctct ttggttttgt ccgggtaggt cgtgagcaat ccggtagtgc 1860 cgatgccaag gctaatcgac gccggacgga ctagactact gtagtagact gtagaggtgt 1920 accgttgcta cttccgtggc gtttgtctgc atgattagga gagaaaactg gcggtggttc 1980 gaggactcta cctgccgatc gagtgagtca agcgagccac ggaaaatgtg taagaaaaaa 2040 atattaagta tgtgtattgt aaaaaaaaaa aaaaaaaaaa aaaaaaaaa 2089 26 539 PRT Triticum aestivum 26 Ala Pro Leu Asn Tyr Thr Gln Gly Gly Pro Arg Arg His Asn Pro Gln 1 5 10 15 Ala Ala Ser Arg Gly Ala Ser Ser Thr Met Asp Arg Ala Ala Leu Pro 20 25 30 Ala Ala Val Glu Pro Lys Lys Lys Gly Asn Val Arg Phe Ala Phe Ala 35 40 45 Cys Ala Ile Leu Ala Ser Met Thr Ser Ile Leu Leu Gly Tyr Asp Ile 50 55 60 Gly Val Met Ser Gly Ala Ser Leu Tyr Ile Gln Lys Asp Leu Lys Ile 65 70 75 80 Asn Asp Thr Gln Leu Glu Val Leu Met Gly Ile Leu Asn Val Tyr Ser 85 90 95 Leu Ile Gly Ser Phe Ala Ala Gly Arg Thr Ser Asp Trp Ile Gly Arg 100 105 110 Arg Phe Thr Ile Val Phe Ala Ala Val Ile Phe Phe Ala Gly Ala Leu 115 120 125 Ile Met Gly Phe Ser Val Asn Tyr Ala Met Leu Met Phe Gly Arg Phe 130 135 140 Val Ala Gly Ile Gly Val Gly Tyr Ala Leu Met Ile Ala Pro Val Asn 145 150 155 160 Thr Gly Glu Val Ser Pro Ala Ser Ala Arg Gly Val Leu Thr Ser Phe 165 170 175 Pro Glu Val Phe Ile Asn Phe Gly Ile Leu Leu Gly Tyr Val Ser Asn 180 185 190 Phe Ala Phe Ala Arg Leu Ser Leu Arg Leu Gly Trp Arg Ile Met Leu 195 200 205 Gly Ile Gly Ala Val Pro Ser Val Leu Leu Ala Phe Met Val Leu Gly 210 215 220 Met Pro Glu Ser Pro Arg Trp Leu Val Met Lys Gly Arg Leu Ala Asp 225 230 235 240 Ala Lys Val Val Leu Ala Lys Thr Ser Asp Thr Pro Glu Glu Ala Ala 245 250 255 Glu Arg Ile Ala Asp Ile Lys Thr Ala Ala Gly Ile Pro Leu Gly Leu 260 265 270 Asp Gly Asp Val Val Pro Val Pro Lys Asn Lys Gly Ser Ser Glu Glu 275 280 285 Lys Arg Val Leu Lys Asp Leu Ile Leu Ser Pro Thr Ile Ala Met Arg 290 295 300 His Ile Leu Ile Ala Gly Ile Gly Ile His Phe Phe Gln Gln Ser Ser 305 310 315 320 Gly Ile Asp Ala Val Val Leu Tyr Ser Pro Leu Val Phe Lys Ser Ala 325 330 335 Gly Ile Thr Gly Asp Ser Arg Leu Arg Gly Thr Thr Val Ala Val Gly 340 345 350 Ala Thr Asn Thr Val Phe Ile Leu Val Ala Thr Phe Leu Leu Asp Arg 355 360 365 Ile Arg Arg Arg Pro Leu Val Leu Thr Ser Thr Gly Gly Met Leu Val 370 375 380 Ser Leu Val Gly Leu Ala Thr Gly Leu Thr Val Ile Ser Arg His Pro 385 390 395 400 Asp Glu Lys Ile Thr Trp Ala Ile Val Leu Cys Ile Phe Cys Ile Met 405 410 415 Ala Tyr Val Ala Phe Phe Ser Ile Gly Leu Gly Pro Ile Thr Trp Val 420 425 430 Tyr Ser Ser Glu Ile Phe Pro Leu His Val Arg Ala Leu Gly Cys Ser 435 440 445 Leu Gly Val Ala Val Asn Arg Leu Thr Ser Gly Val Ile Ser Met Thr 450 455 460 Phe Ile Ser Leu Ser Lys Ala Met Thr Ile Gly Gly Ala Phe Phe Leu 465 470 475 480 Phe Ala Gly Ile Ala Ser Phe Ala Trp Val Phe Phe Phe Ala Tyr Leu 485 490 495 Pro Glu Thr Arg Gly Arg Thr Leu Glu Asp Met Ser Ser Leu Phe Gly 500 505 510 Asn Thr Ala Thr His Lys Gln Gly Ala Ala Glu Ala Asp Asp Asp Ala 515 520 525 Gly Glu Lys Lys Val Glu Met Ala Ala Thr Asn 530 535 27 1872 DNA Triticum aestivum 27 gcacgagctc atcactaggc tgtcagtctg tctgttcaac gaacgatcag ttcgtcctaa 60 gcagatgaaa atgtctccgg aaagaaaagg agcggaggac aaggaagaag gatcgaggat 120 ggcttctgct gcgctcccgg agccgggggc agtccatcca aggaacaagg gcaatttcaa 180 gtacgccttc acctgcgccc tctgtgcttc catggccacc atcgtcctcg gctacgacgt 240 tggggtgatg agcggtgcgt cgctgtacat caagagggac ctgcagatca cggacgtgca 300 gctggagatc atgatgggca tcctgagcgt gtacgcgctc atcgggtcct tcctcggcgc 360 gaggacgtcc gactgggtcg gccgccgcgt caccgtcgtc ttcgcggccg ccatcttcaa 420 caacggctcc ttgctcatgg gcttcgcggt caactacgcc atgctcatgg tcgggcgctt 480 cgtcaccgga atcggcgtgg gctacgccat catggtcgcg ccagtgtaca cgcccgaggt 540 gtccccggcg tcggcccgcg gcttcctcac gtctttcacc gaggtgttca tcaatgtggg 600 catcctcctt ggctacgtct ccaactacgc cttcgcgcgc ctcccgctcc acctcagctg 660 gcgcgtcatg ctcggcatcg gcgccgtccc gtccgccctg cttgcgctca tggtgttcgg 720 catgccggag tctcctcgct ggctcgtcat gaaaggccgc ctcgcggacg ccagggccgt 780 tctggccaag acctccgaca cgccggagga ggccgtggag cgccttgacc agatcaaggc 840 tgccgccggc atccctaggg aacttgacgg cgacgtggtc gtcatgccta agacaaaagg 900 cggccaggag aagcaggtgt ggaaggagct catcttttcg ccgaccccag ccatgcggcg 960 catactgctc gcggcgctcg gcatccattt ctttcagcag gcgacgggct ccgactccgt 1020 cgtgctctat agcccacgcg tgttccagag cgcgggcatc accggcgaca accacctgct 1080 cggcgccaca tgcgccatgg gggtcatgaa gacgctcttc atcctggtgg ccacgttcca 1140 gctcgaccgc gtcggcaggc ggccgctgct gctgaccagc acggccggca tgctcgcctg 1200 tctcatcggc ctcgggacgg gcctcaccgt cgtgggtcgg cacccggacg ccaaggtccc 1260 gtgggccatc ggcctgtgca tcgtgtccat cttggcctac gtgtccttct tctccatcgg 1320 cctcgggccc ctcaccagcg tgtacacctc ggaggtcttc ccactgcggg tgcgcgcgct 1380 gggcttcgcg ctgggcacgt catgcaaccg cgtcaccagc gccgcggtct ccatgtcctt 1440 cctgtccttg tccaaggcca tcaccatcgg cggcagcttc ttcctgtacg ccggcatcgc 1500 ggcgatagga tggattttct tcttcacctt cattccggag acgcgtggcc tgccgctcga 1560 ggagataggg aagcttttcg gcatgacgga cacggccgtc gaagcccaag acaccgccac 1620 gaaagacaag gcgaaagtag gggagatgaa ctagtgagct agacgtcaac caactgttac 1680 cgatgtacta ccatagagat gtatctgatc aacgtggcaa tataagtgtc acggactctt 1740 ggtgctcatt gatggattgt ttggataaaa tttcaagaga attgtttcaa gtttggatcc 1800 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1860 aaaaaaaaaa aa 1872 28 529 PRT Triticum aestivum 28 Met Lys Met Ser Pro Glu Arg Lys Gly Ala Glu Asp Lys Glu Glu Gly 1 5 10 15 Ser Arg Met Ala Ser Ala Ala Leu Pro Glu Pro Gly Ala Val His Pro 20 25 30 Arg Asn Lys Gly Asn Phe Lys Tyr Ala Phe Thr Cys Ala Leu Cys Ala 35 40 45 Ser Met Ala Thr Ile Val Leu Gly Tyr Asp Val Gly Val Met Ser Gly 50 55 60 Ala Ser Leu Tyr Ile Lys Arg Asp Leu Gln Ile Thr Asp Val Gln Leu 65 70 75 80 Glu Ile Met Met Gly Ile Leu Ser Val Tyr Ala Leu Ile Gly Ser Phe 85 90 95 Leu Gly Ala Arg Thr Ser Asp Trp Val Gly Arg Arg Val Thr Val Val 100 105 110 Phe Ala Ala Ala Ile Phe Asn Asn Gly Ser Leu Leu Met Gly Phe Ala 115 120 125 Val Asn Tyr Ala Met Leu Met Val Gly Arg Phe Val Thr Gly Ile Gly 130 135 140 Val Gly Tyr Ala Ile Met Val Ala Pro Val Tyr Thr Pro Glu Val Ser 145 150 155 160 Pro Ala Ser Ala Arg Gly Phe Leu Thr Ser Phe Thr Glu Val Phe Ile 165 170 175 Asn Val Gly Ile Leu Leu Gly Tyr Val Ser Asn Tyr Ala Phe Ala Arg 180 185 190 Leu Pro Leu His Leu Ser Trp Arg Val Met Leu Gly Ile Gly Ala Val 195 200 205 Pro Ser Ala Leu Leu Ala Leu Met Val Phe Gly Met Pro Glu Ser Pro 210 215 220 Arg Trp Leu Val Met Lys Gly Arg Leu Ala Asp Ala Arg Ala Val Leu 225 230 235 240 Ala Lys Thr Ser Asp Thr Pro Glu Glu Ala Val Glu Arg Leu Asp Gln 245 250 255 Ile Lys Ala Ala Ala Gly Ile Pro Arg Glu Leu Asp Gly Asp Val Val 260 265 270 Val Met Pro Lys Thr Lys Gly Gly Gln Glu Lys Gln Val Trp Lys Glu 275 280 285 Leu Ile Phe Ser Pro Thr Pro Ala Met Arg Arg Ile Leu Leu Ala Ala 290 295 300 Leu Gly Ile His Phe Phe Gln Gln Ala Thr Gly Ser Asp Ser Val Val 305 310 315 320 Leu Tyr Ser Pro Arg Val Phe Gln Ser Ala Gly Ile Thr Gly Asp Asn 325 330 335 His Leu Leu Gly Ala Thr Cys Ala Met Gly Val Met Lys Thr Leu Phe 340 345 350 Ile Leu Val Ala Thr Phe Gln Leu Asp Arg Val Gly Arg Arg Pro Leu 355 360 365 Leu Leu Thr Ser Thr Ala Gly Met Leu Ala Cys Leu Ile Gly Leu Gly 370 375 380 Thr Gly Leu Thr Val Val Gly Arg His Pro Asp Ala Lys Val Pro Trp 385 390 395 400 Ala Ile Gly Leu Cys Ile Val Ser Ile Leu Ala Tyr Val Ser Phe Phe 405 410 415 Ser Ile Gly Leu Gly Pro Leu Thr Ser Val Tyr Thr Ser Glu Val Phe 420 425 430 Pro Leu Arg Val Arg Ala Leu Gly Phe Ala Leu Gly Thr Ser Cys Asn 435 440 445 Arg Val Thr Ser Ala Ala Val Ser Met Ser Phe Leu Ser Leu Ser Lys 450 455 460 Ala Ile Thr Ile Gly Gly Ser Phe Phe Leu Tyr Ala Gly Ile Ala Ala 465 470 475 480 Ile Gly Trp Ile Phe Phe Phe Thr Phe Ile Pro Glu Thr Arg Gly Leu 485 490 495 Pro Leu Glu Glu Ile Gly Lys Leu Phe Gly Met Thr Asp Thr Ala Val 500 505 510 Glu Ala Gln Asp Thr Ala Thr Lys Asp Lys Ala Lys Val Gly Glu Met 515 520 525 Asn 29 729 PRT Arabidopsis thaliana 29 Met Ser Gly Ala Val Leu Val Ala Ile Ala Ala Ala Val Gly Asn Leu 1 5 10 15 Leu Gln Gly Trp Asp Asn Ala Thr Ile Ala Gly Ala Val Leu Tyr Ile 20 25 30 Lys Lys Glu Phe Asn Leu Glu Ser Asn Pro Ser Val Glu Gly Leu Ile 35 40 45 Val Ala Met Ser Leu Ile Gly Ala Thr Leu Ile Thr Thr Cys Ser Gly 50 55 60 Gly Val Ala Asp Trp Leu Gly Arg Arg Pro Met Leu Ile Leu Ser Ser 65 70 75 80 Ile Leu Tyr Phe Val Gly Ser Leu Val Met Leu Trp Ser Pro Asn Val 85 90 95 Tyr Val Leu Leu Leu Gly Arg Leu Leu Asp Gly Phe Gly Val Gly Leu 100 105 110 Val Val Thr Leu Val Pro Ile Tyr Ile Ser Glu Thr Ala Pro Pro Glu 115 120 125 Ile Arg Gly Leu Leu Asn Thr Leu Pro Gln Phe Thr Gly Ser Gly Gly 130 135 140 Met Phe Leu Ser Tyr Cys Met Val Phe Gly Met Ser Leu Met Pro Ser 145 150 155 160 Pro Ser Trp Arg Leu Met Leu Gly Val Leu Phe Ile Pro Ser Leu Val 165 170 175 Phe Phe Phe Leu Thr Val Phe Phe Leu Pro Glu Ser Pro Arg Trp Leu 180 185 190 Val Ser Lys Gly Arg Met Leu Glu Ala Lys Arg Val Leu Gln Arg Leu 195 200 205 Arg Gly Arg Glu Asp Val Ser Gly Glu Met Ala Leu Leu Val Glu Gly 210 215 220 Leu Gly Ile Gly Gly Glu Thr Thr Ile Glu Glu Tyr Ile Ile Gly Pro 225 230 235 240 Ala Asp Glu Val Thr Asp Asp His Asp Ile Ala Val Asp Lys Asp Gln 245 250 255 Ile Lys Leu Tyr Gly Ala Glu Glu Gly Leu Ser Trp Val Ala Arg Pro 260 265 270 Val Lys Gly Gly Ser Thr Met Ser Val Leu Ser Arg His Gly Ser Thr 275 280 285 Met Ser Arg Arg Gln Gly Ser Leu Ile Asp Pro Leu Val Thr Leu Phe 290 295 300 Gly Ser Val His Glu Lys Met Pro Asp Thr Gly Ser Met Arg Ser Ala 305 310 315 320 Leu Phe Pro His Phe Gly Ser Met Phe Ser Val Gly Gly Asn Gln Pro 325 330 335 Arg His Glu Asp Trp Asp Glu Glu Asn Leu Val Gly Glu Gly Glu Asp 340 345 350 Tyr Pro Ser Asp His Gly Asp Asp Ser Glu Asp Asp Leu His Ser Pro 355 360 365 Leu Ile Ser Arg Gln Thr Thr Ser Met Glu Lys Asp Met Pro His Thr 370 375 380 Ala His Gly Thr Leu Ser Thr Phe Arg His Gly Ser Gln Val Gln Gly 385 390 395 400 Ala Gln Gly Glu Gly Ala Gly Ser Met Gly Ile Gly Gly Gly Trp Gln 405 410 415 Val Ala Trp Lys Trp Thr Glu Arg Glu Asp Glu Ser Gly Gln Lys Glu 420 425 430 Glu Gly Phe Pro Gly Ser Arg Arg Gly Ser Ile Val Ser Leu Pro Gly 435 440 445 Gly Asp Gly Thr Gly Glu Ala Asp Phe Val Gln Ala Ser Ala Leu Val 450 455 460 Ser Gln Pro Ala Leu Tyr Ser Lys Asp Leu Leu Lys Glu His Thr Ile 465 470 475 480 Gly Pro Ala Met Val His Pro Ser Glu Thr Thr Lys Gly Ser Ile Trp 485 490 495 His Asp Leu His Asp Pro Gly Val Lys Arg Ala Leu Val Val Gly Val 500 505 510 Gly Leu Gln Ile Leu Gln Gln Phe Ser Gly Ile Asn Gly Val Leu Tyr 515 520 525 Tyr Thr Pro Gln Ile Leu Glu Gln Ala Gly Val Gly Ile Leu Leu Ser 530 535 540 Asn Met Gly Ile Ser Ser Ser Ser Ala Ser Leu Leu Ile Ser Ala Leu 545 550 555 560 Thr Thr Phe Val Met Leu Pro Ala Ile Ala Val Ala Met Arg Leu Met 565 570 575 Asp Leu Ser Gly Arg Arg Thr Leu Leu Leu Thr Thr Ile Pro Ile Leu 580 585 590 Ile Ala Ser Leu Leu Val Leu Val Ile Ser Asn Leu Val His Met Asn 595 600 605 Ser Ile Val His Ala Val Leu Ser Thr Val Ser Val Val Leu Tyr Phe 610 615 620 Cys Phe Phe Val Met Gly Phe Gly Pro Ala Pro Asn Ile Leu Cys Ser 625 630 635 640 Glu Ile Phe Pro Thr Arg Val Arg Gly Ile Cys Ile Ala Ile Cys Ala 645 650 655 Leu Thr Phe Trp Ile Cys Asp Ile Ile Val Thr Tyr Ser Leu Pro Val 660 665 670 Leu Leu Lys Ser Ile Gly Leu Ala Gly Val Phe Gly Met Tyr Ala Ile 675 680 685 Val Cys Cys Ile Ser Trp Val Phe Val Phe Ile Lys Val Pro Glu Thr 690 695 700 Lys Gly Met Pro Leu Glu Val Ile Thr Glu Phe Phe Ser Val Gly Ala 705 710 715 720 Arg Gln Ala Glu Ala Ala Lys Asn Glu 725 30 549 PRT Beta vulgaris 30 Met Ser Glu Gly Thr Asn Lys Ala Met Ser Asp Pro Pro Pro Thr Thr 1 5 10 15 Ala Ser Lys Val Ile Ala Asp Phe Asp Pro Leu Lys Lys Pro Pro Lys 20 25 30 Arg Asn Lys Phe Ala Phe Ala Cys Ala Thr Leu Ala Ser Met Thr Ser 35 40 45 Val Leu Leu Gly Tyr Asp Ile Gly Val Met Ser Gly Ala Ile Ile Tyr 50 55 60 Leu Lys Glu Asp Trp His Ile Ser Asp Thr Gln Ile Gly Val Leu Val 65 70 75 80 Gly Ile Leu Asn Ile Tyr Cys Leu Phe Gly Ser Phe Ala Ala Gly Arg 85 90 95 Thr Ser Asp Trp Ile Gly Arg Arg Tyr Thr Ile Val Leu Ala Gly Ala 100 105 110 Ile Phe Phe Val Gly Ala Leu Leu Met Gly Phe Ala Thr Asn Tyr Ala 115 120 125 Phe Leu Met Val Gly Arg Phe Val Thr Gly Ile Gly Val Gly Tyr Ala 130 135 140 Leu Met Ile Ala Pro Val Tyr Thr Ala Glu Val Ser Pro Ala Ser Ser 145 150 155 160 Arg Gly Phe Leu Thr Ser Phe Pro Glu Val Phe Ile Asn Ala Gly Ile 165 170 175 Leu Leu Gly Tyr Ile Ser Asn Leu Ala Phe Ser Ser Leu Pro Thr His 180 185 190 Leu Ser Trp Arg Phe Met Leu Gly Ile Gly Ala Ile Pro Ser Ile Phe 195 200 205 Leu Ala Ile Gly Val Leu Ala Met Pro Glu Ser Pro Arg Trp Leu Val 210 215 220 Met Gln Gly Arg Leu Gly Asp Ala Lys Lys Val Leu Asn Arg Ile Ser 225 230 235 240 Asp Ser Pro Glu Glu Ala Gln Leu Arg Leu Ser Glu Ile Lys Gln Thr 245 250 255 Ala Gly Ile Pro Ala Glu Cys Asp Glu Asp Ile Tyr Lys Val Glu Lys 260 265 270 Thr Lys Ile Lys Ser Gly Asn Ala Val Trp Lys Glu Leu Phe Phe Asn 275 280 285 Pro Thr Pro Ala Val Arg Arg Ala Val Ile Ala Gly Ile Gly Ile His 290 295 300 Phe Phe Gln Gln Ala Ser Gly Ile Asp Ala Val Val Leu Tyr Ser Pro 305 310 315 320 Arg Ile Phe Gln Ser Ala Gly Ile Thr Asn Ala Arg Lys Gln Leu Leu 325 330 335 Ala Thr Val Ala Val Gly Val Val Lys Thr Leu Phe Ile Leu Val Ala 340 345 350 Thr Phe Gln Leu Asp Lys Tyr Gly Arg Arg Pro Leu Leu Leu Thr Ser 355 360 365 Val Gly Gly Met Ile Ile Ala Ile Leu Thr Leu Ala Met Ser Leu Thr 370 375 380 Val Ile Asp His Ser His His Lys Ile Thr Trp Ala Ile Ala Leu Cys 385 390 395 400 Ile Thr Met Val Cys Ala Val Val Ala Ser Phe Ser Ile Gly Leu Gly 405 410 415 Pro Ile Thr Trp Val Tyr Ser Ser Glu Val Phe Pro Leu Arg Leu Arg 420 425 430 Ala Gln Gly Thr Ser Met Gly Val Ala Val Asn Arg Val Val Ser Gly 435 440 445 Val Ile Ser Ile Phe Phe Leu Pro Leu Ser His Lys Ile Thr Thr Gly 450 455 460 Gly Ala Phe Phe Leu Phe Gly Gly Ile Ala Ile Ile Ala Trp Phe Phe 465 470 475 480 Phe Leu Thr Phe Leu Pro Glu Thr Arg Gly Arg Thr Leu Glu Asn Met 485 490 495 His Glu Leu Phe Glu Asp Phe Arg Trp Arg Glu Ser Phe Pro Gly Asn 500 505 510 Lys Ser Asn Asn Asp Glu Asn Ser Thr Arg Lys Gln Ser Asn Gly Asn 515 520 525 Asp Lys Ser Gln Val Gln Leu Gly Glu Thr Thr Thr Ser Thr Thr Val 530 535 540 Thr Asn Asp Asn His 545 

What is claimed is:
 1. An isolated polynucleotide comprising: (a) a first nucleotide sequence encoding a first polypeptide, wherein the first polypeptide comprises at least 117 amino acid residues, wherein the first polypeptide is a sugar transport protein, and wherein the amino acid sequence of the first polypeptide has 90% identity when compared to the amino acid sequence of SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:12, (b) a second nucleotide sequence encoding a second polypeptide, wherein the second polypeptide comprises at least 228 amino acid residues, wherein the second polypeptide is a sugar transport protein, and wherein the amino acid sequence of the second polypeptide has 90% identity when compared to the amino acid sequence of SEQ ID NO:10, SEQ ID NO:14, or SEQ ID NO:16, (c) a third nucleotide sequence encoding a third polypeptide, wherein the third polypeptide comprises at least 737 amino acid residues, wherein the third polypeptide is a sugar transport protein, and wherein the amino acid sequence of the third polypeptide has 90% identity when compared to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8, or (d) a fourth nucleotide sequence comprising the complement of the first, second, or third nucleotide sequence.
 2. The polynucleotide of claim 1, wherein the amino acid sequence of the first polypeptide has 95% identity when compared to the amino acid sequence of SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:12, wherein the amino acid sequence of the second polypeptide has 95% identity when compared to the amino acid sequence of SEQ ID NO:10, SEQ ID NO:14, or SEQ ID NO:16, and wherein the amino acid sequence of the third polypeptide has 95% identity when compared to the amino acid sequence of SEQ ID NO:2 or SEQ ID NO:8.
 3. The polynucleotide of claim 1 comprising the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, or SEQ ID NO:15. 4.The polynucleotide of claim 1 wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, or SEQ ID NO:16.
 5. A chimeric gene comprising the polynucleotide of claim 1 operably linked to a regulatory sequence.
 6. A vector comprising the polynucleotide of claim
 1. 7. A polynucleotide comprising a nucleotide sequence containing at least 30 nucleotides, wherein the nucleotide sequence is comprised by the polynucleotide of claim
 1. 8. A method for transforming a cell comprising transforming a cell with the polynucleotide of claim
 1. 9. A cell comprising the chimeric gene of claim
 5. 10. A method for producing a plant comprising transforming a plant cell with the chimeric gene of claim 5 and regenerating a plant from the transformed plant cell.
 11. A plant comprising the chimeric gene of claim
 5. 12. A seed comprising the chimeric gene of claim
 5. 